From 6ca2eab9493a9d97629e567455ffcba1cd04f544 Mon Sep 17 00:00:00 2001 From: WackerO Date: Tue, 10 Oct 2023 16:15:25 +0200 Subject: [PATCH 01/73] Changed versions to dev --- .github/workflows/ci.yml | 8 ++++---- Dockerfile | 6 +++--- environment.yml | 2 +- modules/local/report.nf | 2 +- nextflow.config | 2 +- 5 files changed, 10 insertions(+), 10 deletions(-) diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index e40f2eb6..d1e1401f 100644 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -40,14 +40,14 @@ jobs: environment.yml - name: Build new docker image if: env.MATCHED_FILES - run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:2.2 + run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:dev # Change the version above and the third version below before/after release - name: Pull docker image if: ${{ !env.MATCHED_FILES }} run: | docker pull ghcr.io/qbic-pipelines/rnadeseq:dev - docker tag ghcr.io/qbic-pipelines/rnadeseq:dev ghcr.io/qbic-pipelines/rnadeseq:2.2 + docker tag ghcr.io/qbic-pipelines/rnadeseq:dev ghcr.io/qbic-pipelines/rnadeseq:dev - name: Install Nextflow uses: nf-core/setup-nextflow@v1 @@ -94,14 +94,14 @@ jobs: environment.yml - name: Build new docker image if: env.MATCHED_FILES - run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:2.2 + run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:dev # Change the version above and the third version below before/after release - name: Pull docker image if: ${{ !env.MATCHED_FILES }} run: | docker pull ghcr.io/qbic-pipelines/rnadeseq:dev - docker tag ghcr.io/qbic-pipelines/rnadeseq:dev ghcr.io/qbic-pipelines/rnadeseq:2.2 + docker tag ghcr.io/qbic-pipelines/rnadeseq:dev ghcr.io/qbic-pipelines/rnadeseq:dev - name: Install Nextflow uses: nf-core/setup-nextflow@v1 diff --git a/Dockerfile b/Dockerfile index c13267b3..4f8ce7fe 100644 --- a/Dockerfile +++ b/Dockerfile @@ -5,14 +5,14 @@ LABEL org.opencontainers.image.authors="Gisela Gabernet, Alexander Peltzer, Oska LABEL org.opencontainers.image.licenses=MIT COPY environment.yml / #RUN conda install -c conda-forge mamba -RUN mamba env create --file /environment.yml -p /opt/conda/envs/qbic-pipelines-rnadeseq-2.2 && \ +RUN mamba env create --file /environment.yml -p /opt/conda/envs/qbic-pipelines-rnadeseq-dev && \ mamba clean --all --yes RUN apt-get update -qq && \ apt-get install -y zip procps ghostscript # Add conda installation dir to PATH -ENV PATH /opt/conda/envs/qbic-pipelines-rnadeseq-2.2/bin:$PATH +ENV PATH /opt/conda/envs/qbic-pipelines-rnadeseq-dev/bin:$PATH # Dump the details of the installed packates to a file for posterity -RUN mamba env export --name qbic-pipelines-rnadeseq-2.2 > qbic-pipelines-rnadeseq-2.2.yml +RUN mamba env export --name qbic-pipelines-rnadeseq-dev > qbic-pipelines-rnadeseq-dev.yml # Instruct R processes to use these empty files instead of clashing with a local config RUN touch .Rprofile RUN touch .Renviron diff --git a/environment.yml b/environment.yml index cf3dc64d..cede896c 100644 --- a/environment.yml +++ b/environment.yml @@ -1,7 +1,7 @@ # You can use this file to create a conda environment for this pipeline: # conda env create -f environment.yml # use this to find packages: https://anaconda.org/ -name: qbic-pipelines-rnadeseq-2.2 +name: qbic-pipelines-rnadeseq-dev channels: - bioconda - conda-forge diff --git a/modules/local/report.nf b/modules/local/report.nf index eac509d0..167fceb5 100644 --- a/modules/local/report.nf +++ b/modules/local/report.nf @@ -1,6 +1,6 @@ process REPORT { - container 'ghcr.io/qbic-pipelines/rnadeseq:2.2' + container 'ghcr.io/qbic-pipelines/rnadeseq:dev' input: path gene_counts diff --git a/nextflow.config b/nextflow.config index 9018f037..21173295 100644 --- a/nextflow.config +++ b/nextflow.config @@ -252,7 +252,7 @@ manifest { description = """Differential gene expression analysis and pathway analysis of RNAseq data""" mainScript = 'main.nf' nextflowVersion = '!>=23.04.0' - version = '2.2' + version = '2.3dev' doi = '' } From 2084b1365b2312b401093956b2e6720341ae5f76 Mon Sep 17 00:00:00 2001 From: WackerO Date: Tue, 10 Oct 2023 16:16:53 +0200 Subject: [PATCH 02/73] Bugfix for nonconformable arrays bug --- assets/RNAseq_report.Rmd | 5 +++++ 1 file changed, 5 insertions(+) diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index b1aa99b8..0c69f405 100644 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -1437,6 +1437,11 @@ write(contrast_names, file="contrast_names.txt", sep="\t") # Remove identical columns of DE_genes_df DE_genes_df$DE_genes_df <- NULL idx <- duplicated(t(DE_genes_df)) + +# If any padj or logFC columns were marked for removal (TRUE), undo that by setting FALSE as we need these cols +idx[grepl("padj", as.character(names(idx)))] <- FALSE +idx[grepl("log2FoldChange", as.character(names(idx)))] <- FALSE + DE_genes_df <- DE_genes_df[, !idx] DE_genes_df$Ensembl_ID <- row.names(DE_genes_df) DE_genes_df <- DE_genes_df[,c(dim(DE_genes_df)[2],1:dim(DE_genes_df)[2]-1)] From ef825c281bb7c266e57f5fcd79cd4ccee7bce357 Mon Sep 17 00:00:00 2001 From: WackerO Date: Thu, 12 Oct 2023 15:55:24 +0200 Subject: [PATCH 03/73] also added padj to hovertext in volcano --- assets/RNAseq_report.Rmd | 8 ++++++-- 1 file changed, 6 insertions(+), 2 deletions(-) diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index 0c69f405..92a4a1b6 100644 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -1557,17 +1557,21 @@ for (file in allgenes_files){ } DE_all <- ldply(table_list, rbind) DE_all$logpval <- -log10(DE_all$padj) +DE_all$truelogpval <- DE_all$logpval # Save true value for hovertext DE_all$logpval[DE_all$logpval > 16] <- 17 DE_all <- na.omit(DE_all) +DE_all <- DE_all %>% mutate(color = ifelse(abs(DE_all$log2FoldChange) >= params$logFC_threshold & DE_all$logpval >= -log10(params$adj_pval_threshold), "red", "black")) # Create column which assigns each entry a plot color so that coloring is consistent even when no DE genes are found log2FoldChange_min <- min(DE_all$log2FoldChange) log2FoldChange_max <- max(DE_all$log2FoldChange) # get quotient of log2FoldChange range and 10 to determine if xticks of plot need to be scaled discretely (otherwise, for large FC ranges, there will be too many xticks and they will overlap) log2FoldChange_quotient <- length(seq(log2FoldChange_min, log2FoldChange_max, 10)) -pg <- ggplot(DE_all, aes(x=log2FoldChange, y=logpval, text=paste("Gene: ", gene_name, "
", "Log2FC: ", formatC(log2FoldChange, digits=2)))) + +pg <- ggplot(DE_all, aes(x=log2FoldChange, y=logpval, text=paste("Gene: ", gene_name, + "
", "Log2FC: ", formatC(log2FoldChange, digits=2), + "
", "-Log10padj: ", formatC(truelogpval, digits=2)))) + #this goes after alpha=0.5: - geom_jitter(alpha=0.5, width = 0.2, aes(color=ifelse(abs(DE_all$log2FoldChange) >= params$logFC_threshold & DE_all$logpval >= -log10(params$adj_pval_threshold), "Differentially expressed genes", "Non-differentially expressed genes"))) + + geom_jitter(alpha=0.5, width = 0.2, aes(color=ifelse(DE_all$color == "red", "Differentially expressed genes", "Non-differentially expressed genes"))) + geom_hline(yintercept = 16, linetype= "dashed", size = 0.2, color = "grey") + geom_hline(yintercept = -log10(params$adj_pval_threshold), size = 0.2, color = "grey") + geom_vline(xintercept = -params$logFC_threshold, size = 0.2, color = "grey") + From b735596375d9275fa7920a4ffd3982e3b14cd8e8 Mon Sep 17 00:00:00 2001 From: WackerO Date: Fri, 13 Oct 2023 09:26:04 +0200 Subject: [PATCH 04/73] Updated changelog --- CHANGELOG.md | 11 +++++++++++ 1 file changed, 11 insertions(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index 56d1d982..2f033550 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -3,6 +3,17 @@ The format is based on [Keep a Changelog](https://keepachangelog.com/en/1.0.0/) and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0.html). +## 2.3dev + +### Added + +- [#221](https://github.com/qbic-pipelines/rnadeseq/pull/221) Add padj to volcano hovertext + +### Fixed + +- [#221](https://github.com/qbic-pipelines/rnadeseq/pull/221) Fix non-conformable arrays bug, fix wrong volcano colors when no DE genes + + ## 2.2 Avenue of Poplars ### Added From 63700be470db0aea61182c1a598fd1fe3962d104 Mon Sep 17 00:00:00 2001 From: WackerO Date: Fri, 13 Oct 2023 09:28:06 +0200 Subject: [PATCH 05/73] linting --- lib/NfcoreTemplate.groovy | 16 ++++++++++++++++ 1 file changed, 16 insertions(+) diff --git a/lib/NfcoreTemplate.groovy b/lib/NfcoreTemplate.groovy index 000bb929..c084c800 100755 --- a/lib/NfcoreTemplate.groovy +++ b/lib/NfcoreTemplate.groovy @@ -3,6 +3,7 @@ // import org.yaml.snakeyaml.Yaml +import groovy.json.JsonOutput class NfcoreTemplate { @@ -222,6 +223,21 @@ class NfcoreTemplate { } } + // + // Dump pipeline parameters in a json file + // + public static void dump_parameters(workflow, params) { + def output_d = new File("${params.outdir}/pipeline_info/") + if (!output_d.exists()) { + output_d.mkdirs() + } + + def timestamp = new java.util.Date().format( 'yyyy-MM-dd_HH-mm-ss') + def output_pf = new File(output_d, "params_${timestamp}.json") + def jsonStr = JsonOutput.toJson(params) + output_pf.text = JsonOutput.prettyPrint(jsonStr) + } + // // Print pipeline summary on completion // From ef2f16f66d8af39fe6ccb3627199fad18970f025 Mon Sep 17 00:00:00 2001 From: WackerO Date: Fri, 13 Oct 2023 09:29:20 +0200 Subject: [PATCH 06/73] prettier --- CHANGELOG.md | 1 - 1 file changed, 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 2f033550..66a9e5c2 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -13,7 +13,6 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 - [#221](https://github.com/qbic-pipelines/rnadeseq/pull/221) Fix non-conformable arrays bug, fix wrong volcano colors when no DE genes - ## 2.2 Avenue of Poplars ### Added From e133651a9c9cc2e27ad9fe83aa995b6b14996a2c Mon Sep 17 00:00:00 2001 From: WackerO Date: Fri, 13 Oct 2023 14:38:35 +0200 Subject: [PATCH 07/73] fixed tests --- tests/test.yml | 19 ++++++++++--------- tests/test_full.yml | 18 ++++++++++++------ tests/test_star_rsem.yml | 8 +++----- tests/test_star_salmon.yml | 2 +- 4 files changed, 26 insertions(+), 21 deletions(-) diff --git a/tests/test.yml b/tests/test.yml index 56ff217d..401899c1 100644 --- a/tests/test.yml +++ b/tests/test.yml @@ -66,14 +66,12 @@ - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.pdf - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_pathway_enrichment_results.tsv - md5sum: be0f292a205f2ecd453ee27ecdc62615 + contains: ["Complement and coagulation cascades", "Muscle contraction", "Hemostasis"] - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_results.tsv - md5sum: f0e6d8117e3ff05ddabc3abb98f514b7 - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_results.tsv - md5sum: 3d7616fd0f6cb3aaf9f08252b2bcd223 - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_plot.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_plot.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_plot.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04360.DE_contrast_condition_genotype_WT_vs_KO.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04360.png @@ -82,13 +80,16 @@ - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04610.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04610.xml - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_pathway_enrichment_results.tsv - md5sum: 0bd6dfd32bad6f5292f6cde91f019844 + contains: ["IL-17 signaling pathway", "RAF-independent MAPK1/3 activation"] - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_results.tsv - md5sum: 452ca5dd8d5f190648988502f85d3ef1 - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.png - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.svg + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_KEGG_pathway_enrichment_results.tsv + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_results.tsv + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_KEGG_pathway_enrichment_plot.pdf - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_KEGG_pathway_enrichment_plot.png - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_plot.png - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.png + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.svg - path: results_test/RNAseq_report.html diff --git a/tests/test_full.yml b/tests/test_full.yml index c4477cbf..557410cc 100644 --- a/tests/test_full.yml +++ b/tests/test_full.yml @@ -60,26 +60,32 @@ - path: results_test/differential_gene_expression/plots/PCA_plot.pdf - path: results_test/differential_gene_expression/plots/PCA_plot.png - path: results_test/differential_gene_expression/plots/PCA_plot.svg + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_venn_diagram.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_venn_diagram.png + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_venn_diagram.svg - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.pdf - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_pathway_enrichment_results.tsv - md5sum: be0f292a205f2ecd453ee27ecdc62615 + contains: ["Complement and coagulation cascades", "Muscle contraction", "Hemostasis"] - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_results.tsv - md5sum: f0e6d8117e3ff05ddabc3abb98f514b7 - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_results.tsv - md5sum: 3d7616fd0f6cb3aaf9f08252b2bcd223 - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_plot.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_plot.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_plot.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04360.DE_contrast_condition_genotype_WT_vs_KO.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04360.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04360.xml - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_pathway_enrichment_results.tsv - md5sum: 0bd6dfd32bad6f5292f6cde91f019844 + contains: ["IL-17 signaling pathway", "RAF-independent MAPK1/3 activation"] + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_KEGG_pathway_enrichment_results.tsv - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_results.tsv - md5sum: 452ca5dd8d5f190648988502f85d3ef1 + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_KEGG_pathway_enrichment_plot.pdf - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_KEGG_pathway_enrichment_plot.png - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_plot.png - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.png + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.svg - path: results_test/RNAseq_report.html diff --git a/tests/test_star_rsem.yml b/tests/test_star_rsem.yml index c9cbdd49..1cf8eea5 100644 --- a/tests/test_star_rsem.yml +++ b/tests/test_star_rsem.yml @@ -88,11 +88,11 @@ - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_REAC_pathway_enrichment_plot.pdf - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_REAC_pathway_enrichment_plot.png - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_pathway_enrichment_results.tsv - md5sum: b1c0a1374c963fff71083a9cdefe8e9b + contains: ["Cell adhesion molecules", "Cytokine-cytokine receptor interaction", "Amoebiasis"] - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_pathway_enrichment_results.tsv - md5sum: 631e89b4de9042936fa2652bde036e7d + contains: ["Cell adhesion molecules", "Cytokine-cytokine receptor interaction", "Amoebiasis"] - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_pathway_enrichment_results.tsv - md5sum: 80123c4ba2c97eaa0217816f41122855 + contains: ["Cell adhesion molecules", "Cytokine-cytokine receptor interaction", "Amoebiasis"] - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_KEGG_pathway_enrichment_plot.pdf - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_KEGG_pathway_enrichment_plot.png - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_KEGG_pathway_enrichment_plot.pdf @@ -106,7 +106,5 @@ - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_REAC_pathway_enrichment_plot.pdf - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_REAC_pathway_enrichment_plot.png - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_KEGG_pathway_enrichment_results.tsv - md5sum: 276e871c81e85217bbd7eef951676084 - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_REAC_pathway_enrichment_results.tsv - md5sum: 9e6847fe5d631f09305dfa00dd3472d1 - path: results_test/RNAseq_report.html diff --git a/tests/test_star_salmon.yml b/tests/test_star_salmon.yml index dbe49c9a..43b062a3 100644 --- a/tests/test_star_salmon.yml +++ b/tests/test_star_salmon.yml @@ -70,7 +70,7 @@ - path: results_test/differential_gene_expression/plots/PCA_plot.png - path: results_test/differential_gene_expression/plots/PCA_plot.svg - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_pathway_enrichment_results.tsv - md5sum: cd967cc355045d9dc8c0bb6ffcf40b61 + contains: ["Extracellular matrix organization"] - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_pathway_enrichment_results.tsv md5sum: 68b329da9893e34099c7d8ad5cb9c940 - path: results_test/RNAseq_report.html From 21cfa7caa72458124e5140d9aceff73c92fd9845 Mon Sep 17 00:00:00 2001 From: WackerO Date: Tue, 14 Nov 2023 15:36:11 +0100 Subject: [PATCH 08/73] Bugfix for non-conformable arrays when certain contrasts in a dataset have no significant results --- assets/RNAseq_report.Rmd | 13 +++++++++++++ 1 file changed, 13 insertions(+) diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index 92a4a1b6..2b3cb381 100644 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -693,6 +693,7 @@ if (run_rlog){ # rlog transformation rld <- rlog(cds, blind=FALSE) rld_names <- merge(x=gene_names, y=assay(rld), by.x = "Ensembl_ID", by.y="row.names") + rld_names <- rld_names[order(rld_names$Ensembl_ID),] write.table(rld_names, "differential_gene_expression/gene_counts_tables/rlog_transformed_gene_counts.tsv", append = FALSE, quote = FALSE, sep = "\t",eol = "\n", na = "NA", dec = ".", row.names = F, qmethod = c("escape", "double")) # save table to another variable if pathway analysis @@ -706,6 +707,7 @@ if (run_rlog){ # vst transformation vsd <- vst(cds, blind=FALSE, nsub = params$nsub_genes) vsd_names <- merge(x=gene_names, y=assay(vsd), by.x = "Ensembl_ID", by.y="row.names") + vsd_names <- vsd_names[order(vsd_names$Ensembl_ID),] write.table(vsd_names, "differential_gene_expression/gene_counts_tables/vst_transformed_gene_counts.tsv", append = FALSE, quote = FALSE, sep = "\t",eol = "\n", na = "NA", dec = ".", row.names = F, qmethod = c("escape", "double")) # save table to another variable if pathway analysis @@ -1113,9 +1115,20 @@ A PCA of the batch-effect corrected data is shown below and can be found [here]( if (run_rlog) { assay(rld) <- limma::removeBatchEffect(assay(rld), rld$batch) rld_corrected <- rld + out_rld <- as.data.frame(assay(rld_corrected)) + out_rld <- out_rld[order(rownames(out_rld)),] + out_rld <- cbind(Ensembl_ID = rownames(out_rld), gene_name = rld_names$gene_name, out_rld[,!names(out_rld) %in% c("gene_name")]) + + write.table(out_rld, file="differential_gene_expression/gene_counts_tables/rlog_transformed_gene_counts_batchcorrected.tsv", quote=F, sep="\t", row.names=F) } else { assay(vsd) <- limma::removeBatchEffect(assay(vsd), vsd$batch) vsd_corrected <- vsd + + out_vsd <- as.data.frame(assay(vsd_corrected)) + out_vsd <- out_vsd[order(rownames(out_vsd)),] + out_vsd <- cbind(Ensembl_ID = rownames(out_vsd), gene_name = vsd_names$gene_name, out_vsd[,!names(out_vsd) %in% c("gene_name")]) + + write.table(out_vsd, file="differential_gene_expression/gene_counts_tables/vst_transformed_gene_counts_batchcorrected.tsv", quote=F, sep="\t", row.names=F) } pcaData2 <- plotPCA(if (run_rlog) rld else vsd, intgroup=c("combfactor"), ntop = dim(if (run_rlog) rld else vsd)[1], returnData=TRUE) percentVar <- round(100*attr(pcaData, "percentVar")) From 6d26b45601556adb0e168c650103d33ee568d19b Mon Sep 17 00:00:00 2001 From: WackerO Date: Mon, 5 Feb 2024 10:03:13 +0100 Subject: [PATCH 09/73] This should fix the too many devices error --- .devcontainer/devcontainer.json | 0 .editorconfig | 0 .gitattributes | 0 .github/.dockstore.yml | 0 .github/CONTRIBUTING.md | 0 .github/ISSUE_TEMPLATE/bug_report.yml | 0 .github/ISSUE_TEMPLATE/feature_request.yml | 0 .github/PULL_REQUEST_TEMPLATE.md | 0 .github/workflows/awsfulltest.yml | 0 .github/workflows/awstest.yml | 0 .github/workflows/branch.yml | 0 .github/workflows/ci.yml | 0 .github/workflows/clean-up.yml | 0 .github/workflows/fix-linting.yml | 0 .github/workflows/linting.yml | 0 .github/workflows/linting_comment.yml | 0 .github/workflows/push_github_container_reg.yml | 0 .gitignore | 0 .gitpod.yml | 0 .nf-core.yml | 0 .pre-commit-config.yaml | 0 .prettierignore | 0 .prettierrc.yml | 0 CHANGELOG.md | 0 CITATIONS.md | 0 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b/.gitpod.yml old mode 100644 new mode 100755 diff --git a/.nf-core.yml b/.nf-core.yml old mode 100644 new mode 100755 diff --git a/.pre-commit-config.yaml b/.pre-commit-config.yaml old mode 100644 new mode 100755 diff --git a/.prettierignore b/.prettierignore old mode 100644 new mode 100755 diff --git a/.prettierrc.yml b/.prettierrc.yml old mode 100644 new mode 100755 diff --git a/CHANGELOG.md b/CHANGELOG.md old mode 100644 new mode 100755 diff --git a/CITATIONS.md b/CITATIONS.md old mode 100644 new mode 100755 diff --git a/CODE_OF_CONDUCT.md b/CODE_OF_CONDUCT.md old mode 100644 new mode 100755 diff --git a/Dockerfile b/Dockerfile old mode 100644 new mode 100755 diff --git a/LICENSE b/LICENSE old mode 100644 new mode 100755 diff --git a/README.md b/README.md old mode 100644 new mode 100755 diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd old mode 100644 new mode 100755 index b1aa99b8..3f764a9b --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -2032,30 +2032,33 @@ for (file in contrast_files){ tryCatch( { png(filename = paste0("pathway_analysis", "/",fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_",fname, ".png"), width = ifelse(nrow(mat)<=20, 200, 125)+ncol(mat)*40, height = 50+nrow(mat)*50) - pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T) dev.off() + pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T) }, error=function(e) { + dev.off() print(paste0("Could not save Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_",fname, ".png because of the following error:\n", e)) } ) tryCatch( { pdf(paste0("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".pdf"), width = ifelse(nrow(mat)<=20, 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005) - pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T) dev.off() + pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T) }, error=function(e) { + dev.off() print(paste0("Could not save Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".pdf because of the following error:\n", e)) } ) tryCatch( { svg(paste0("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".svg"), width = ifelse(nrow(mat)<=20, 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005) - pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T) dev.off() + pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T) }, error=function(e) { + dev.off() print(paste0("Could not save Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".svg because of the following error:\n", e)) } ) diff --git a/assets/adaptivecard.json b/assets/adaptivecard.json old mode 100644 new mode 100755 diff --git a/assets/email_template.html 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b/tower.yml old mode 100644 new mode 100755 diff --git a/workflows/rnadeseq.nf b/workflows/rnadeseq.nf old mode 100644 new mode 100755 From 9dcd79c813e1589ab4a74d792d84cf4d1c3ef718 Mon Sep 17 00:00:00 2001 From: WackerO Date: Mon, 5 Feb 2024 12:42:52 +0100 Subject: [PATCH 10/73] updated changelog --- CHANGELOG.md | 1 + 1 file changed, 1 insertion(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index 56d1d982..7ada1e03 100755 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -34,6 +34,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### Fixed +- [#222](https://github.com/qbic-pipelines/rnadeseq/pull/222) Fixed too many devices error from tryCatch around normalized heatmaps - [#212](https://github.com/qbic-pipelines/rnadeseq/pull/212) Fixed movability of interactive gostplots - [#208](https://github.com/qbic-pipelines/rnadeseq/pull/208) Fixed relevel bug, the function should now finally work! - [#207](https://github.com/qbic-pipelines/rnadeseq/pull/207) Fixed check of samples in counts vs metadata From 58865a862055f02e9132db0acc54cc3d3510bc6c Mon Sep 17 00:00:00 2001 From: WackerO Date: Mon, 12 Feb 2024 14:48:02 +0100 Subject: [PATCH 11/73] Replaced double dev.offs with finally --- assets/RNAseq_report.Rmd | 25 ++++++++++++++----------- 1 file changed, 14 insertions(+), 11 deletions(-) diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index 3f764a9b..95431f82 100755 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -599,7 +599,8 @@ if (params$input_type %in% c("featurecounts", "smrnaseq")) { } } missing_files <- paste0(substring(missing_files, 1, nchar(missing_files)-2), ".") - stop(missing_files, call.=F) } + stop(missing_files, call.=F) + } #The following steps are necessary for the processing of salmon output as the files do # not contain integer counts and can therefore not be directly used for cds @@ -719,7 +720,7 @@ if (run_rlog){ # Cooks distances: get important for example when checking knock-out and overexpression studies pdf("differential_gene_expression/plots/further_diagnostics_plots/Cooks-distances.pdf") par(mar=c(10,3,3,3)) -par( mfrow = c(1,2)) +par(mfrow = c(1,2)) boxplot(log10(assays(cds)[["cooks"]]), range=0, las=2,ylim = c(-15, 15),main="log10-Cooks") boxplot(log2(assays(cds)[["cooks"]]), range=0, las=2,ylim = c(-15, 15),main="log2-Cooks") dev.off() @@ -1348,6 +1349,7 @@ if (isProvided(params$path_contrast_list)) { contrast_names <- append(contrast_names, contname) } } + if (isProvided(params$path_contrast_pairs)) { contrasts <- read.table(path_contrast_pairs, sep="\t", header = T, colClasses = "character") write.table(contrasts, file="differential_gene_expression/metadata/contrast_pairs.tsv", sep="\t", quote=F, col.names = T, row.names = F) @@ -2027,39 +2029,42 @@ for (file in contrast_files){ # Skip the heatmaps for CI tests because especially png() works only very unreliably for the heatmaps for some reason if (!isProvided(params$citest) & nrow(mat)>1){ dir.create(paste("pathway_analysis", fname, "pathway_heatmaps", sep="/")) - + # Because the image devices sometimes don't work here, surround them with a tryCatch so that the code continues even if a certain heatmap is not saved tryCatch( { png(filename = paste0("pathway_analysis", "/",fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_",fname, ".png"), width = ifelse(nrow(mat)<=20, 200, 125)+ncol(mat)*40, height = 50+nrow(mat)*50) - dev.off() pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T) }, error=function(e) { - dev.off() print(paste0("Could not save Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_",fname, ".png because of the following error:\n", e)) + }, + finally={ + dev.off() } ) tryCatch( { pdf(paste0("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".pdf"), width = ifelse(nrow(mat)<=20, 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005) - dev.off() pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T) }, error=function(e) { - dev.off() print(paste0("Could not save Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".pdf because of the following error:\n", e)) + }, + finally={ + dev.off() } ) tryCatch( { svg(paste0("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".svg"), width = ifelse(nrow(mat)<=20, 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005) - dev.off() pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T) }, error=function(e) { - dev.off() print(paste0("Could not save Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".svg because of the following error:\n", e)) + }, + finally={ + dev.off() } ) } @@ -2073,9 +2078,7 @@ for (file in contrast_files){ print_string <- paste0("Plotting pathway: ", pathway_kegg) gene.data = DE_genes gene.data.subset = gene.data[gene.data$Ensembl_ID %in% gene_list, c("Ensembl_ID","log2FoldChange")] - entrez_ids = AnnotationDbi::mapIds(species_library_installed, keys=as.character(gene.data.subset$Ensembl_ID), column = "ENTREZID", keytype=params$keytype, multiVals="first") - gene.data.subset <- gene.data.subset[!(is.na(entrez_ids)),] if (length(entrez_ids)!=length(unique(entrez_ids))) { From 84a9b9da2017aaae2e9682f1fc4c8fd61ae8aeb0 Mon Sep 17 00:00:00 2001 From: WackerO Date: Thu, 15 Feb 2024 11:16:29 +0100 Subject: [PATCH 12/73] Shortened some output filepaths, added datasources param to make pathway analysis adjustable --- assets/RNAseq_report.Rmd | 67 +++++++++++++++++++++++++++------------- bin/Execute_report.R | 2 ++ conf/test.config | 1 + modules/local/report.nf | 2 ++ nextflow.config | 1 + nextflow_schema.json | 6 +++- 6 files changed, 57 insertions(+), 22 deletions(-) diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index 95431f82..87f09691 100755 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -43,6 +43,7 @@ params: custom_gmt: '' set_background: '' custom_background: '' + datasources: '' #Additional args for the report path_proj_summary: '' @@ -1773,8 +1774,12 @@ BiocManager::install(params$species_library, lib=species_dir, version="3.17", fo library(params$species_library, lib.loc=species_dir, character.only=T) species_library_installed <- get(params$species_library) -# gprofiler pathway / term sources parameters -datasources <- c("KEGG", "REAC") +# If provided, prepare datasources from comma-separated string +if (isProvided(params$datasources)) { + datasources <- unlist(strsplit(params$datasources, ',')) +} else { + datasources <- c() +} # Default to 1 for the nb of genes DE in a pathway min_DEG_pathway <- as.integer(params$min_DEG_pathway) @@ -1804,7 +1809,18 @@ mv_status <- F if (isProvided(params$custom_gmt)){ # If custom GMT file was provided, filter it before uploading to use in gost queries (gprofiler will NOT filter itself, so otherwise the output will contain all kinds of datasources, not just the selected ones) - out_gmt <- (Filter(function(line) any(startsWith(line, datasources)), readLines(params$custom_gmt))) + out_gmt <- readLines(params$custom_gmt) + + # If datasources are also provided, filter the GMT + if (isProvided(params$datasources)) { + go_positions <- grep("^GO", datasources) + if (length(go_positions)) { + datasources <- my_vector[-go_positions] # If there are any datasources starting with GO, remove them (gets rid of GO:BP etc.) + datasources <- append(datasources, "GO") # Then add the single entry GO, as otherwise, the filtering might not recognize these datasources + } + out_gmt <- (Filter(function(line) any(startsWith(line, datasources)), out_gmt)) # Remove GMT lines that don't start with a datasource + } + out_path <- paste0(tools::file_path_sans_ext(basename(params$custom_gmt)), "_filtered.gmt") writeLines(out_gmt, out_path) gost_id <- upload_GMT_file(out_path) @@ -1940,13 +1956,13 @@ for (file in contrast_files){ # Generate non-interactive pathway dotplots in the folder pg <- gostplot(gostres, capped=T, interactive=F) - ggsave(pg, filename = paste0("pathway_analysis", "/", fname, "_gost_pathway_enrichment_plot.pdf"), + ggsave(pg, filename = "pathway_analysis/gost_pathway_enrichment_plot.pdf", device="pdf", height=10, width=15, units="cm", limitsize=F) - ggsave(pg, filename = paste0("pathway_analysis", "/", fname, "_gost_pathway_enrichment_plot.png"), + ggsave(pg, filename = "pathway_analysis/gost_pathway_enrichment_plot.png", device="png", height=10, width=15, units="cm", dpi=300, limitsize=F) - ggsave(pg, filename = paste0("pathway_analysis", "/", fname, "_gost_pathway_enrichment_plot.svg"), + ggsave(pg, filename = "pathway_analysis/gost_pathway_enrichment_plot.svg", device="svg", height=10, width=15, units="cm", dpi=300, limitsize=F) @@ -1972,7 +1988,7 @@ for (file in contrast_files){ # Save pathway enrichment table in tsv format write.table(pathway_gostres_table, - file = paste0("pathway_analysis", "/", fname, "/", fname, "_pathway_enrichment_results.tsv"), + file = paste0("pathway_analysis", "/", fname, "/", "pathway_enrichment_results.tsv"), sep="\t", quote = F, col.names = T, row.names = F) print("------------------------------------") @@ -1994,7 +2010,7 @@ for (file in contrast_files){ # Plotting results for df df_subset <- data.frame(Pathway_name = df$short_name, Pathway_code = df$term_id, DE_genes = df$intersection_size, Pathway_size = df$term_size, Fraction_DE = (df$intersection_size / df$term_size), Padj = df$p_value, DE_genes_names = df$intersection) write.table(df_subset, - file = paste0("pathway_analysis", "/", fname, "/", fname, "_", db_source, "_pathway_enrichment_results.tsv"), + file = paste0("pathway_analysis", "/", fname, "/", db_source, "_pathway_enrichment_results.tsv"), sep="\t", quote = F, col.names = T, row.names = F) # Enriched pathways horizontal barplots of padj values @@ -2006,9 +2022,9 @@ for (file in contrast_files){ scale_fill_continuous(high = "#132B43", low = "#56B1F7") + ggtitle("Enriched pathways") + xlab("") + ylab("Gene fraction (DE genes / Pathway size)") - ggsave(p, filename = paste0("pathway_analysis", "/", fname, "/", fname, "_", db_source, "_pathway_enrichment_plot.pdf"), device = "pdf", height = 5+0.5*nrow(df_subset), units = "cm", limitsize=F) - ggsave(p, filename = paste0("pathway_analysis", "/", fname, "/", fname,"_", db_source, "_pathway_enrichment_plot.png"), device = "png", height = 5+0.5*nrow(df_subset), units = "cm", dpi = 300, limitsize=F) - ggsave(p, filename = paste0("pathway_analysis", "/", fname, "/", fname,"_", db_source, "_pathway_enrichment_plot.svg"), device = "svg", height = 5+0.5*nrow(df_subset), units = "cm", dpi = 300, limitsize=F) + ggsave(p, filename = paste0("pathway_analysis", "/", fname, "/", db_source, "_pathway_enrichment_plot.pdf"), device = "pdf", height = 5+0.5*nrow(df_subset), units = "cm", limitsize=F) + ggsave(p, filename = paste0("pathway_analysis", "/", fname, "/", db_source, "_pathway_enrichment_plot.png"), device = "png", height = 5+0.5*nrow(df_subset), units = "cm", dpi = 300, limitsize=F) + ggsave(p, filename = paste0("pathway_analysis", "/", fname, "/", db_source, "_pathway_enrichment_plot.svg"), device = "svg", height = 5+0.5*nrow(df_subset), units = "cm", dpi = 300, limitsize=F) # Plotting heatmaps and KEGG pathways for all pathways print("Plotting heatmaps...") @@ -2033,11 +2049,11 @@ for (file in contrast_files){ # Because the image devices sometimes don't work here, surround them with a tryCatch so that the code continues even if a certain heatmap is not saved tryCatch( { - png(filename = paste0("pathway_analysis", "/",fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_",fname, ".png"), width = ifelse(nrow(mat)<=20, 200, 125)+ncol(mat)*40, height = 50+nrow(mat)*50) + png(filename = paste0("pathway_analysis", "/",fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, ".png"), width = ifelse(nrow(mat)<=20, 200, 125)+ncol(mat)*40, height = 50+nrow(mat)*50) pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T) }, error=function(e) { - print(paste0("Could not save Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_",fname, ".png because of the following error:\n", e)) + print(paste0("Could not save pathway_analysis", "/",fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, ".png because of the following error:\n", e)) }, finally={ dev.off() @@ -2045,11 +2061,11 @@ for (file in contrast_files){ ) tryCatch( { - pdf(paste0("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".pdf"), width = ifelse(nrow(mat)<=20, 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005) + pdf(paste0("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, ".pdf"), width = ifelse(nrow(mat)<=20, 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005) pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T) }, error=function(e) { - print(paste0("Could not save Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".pdf because of the following error:\n", e)) + print(paste0("Could not save pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, ".pdf because of the following error:\n", e)) }, finally={ dev.off() @@ -2057,11 +2073,11 @@ for (file in contrast_files){ ) tryCatch( { - svg(paste0("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".svg"), width = ifelse(nrow(mat)<=20, 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005) + svg(paste0("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, ".svg"), width = ifelse(nrow(mat)<=20, 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005) pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T) }, error=function(e) { - print(paste0("Could not save Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".svg because of the following error:\n", e)) + print(paste0("Could not save pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, ".svg because of the following error:\n", e)) }, finally={ dev.off() @@ -2440,16 +2456,25 @@ name_species = params$species_library version_species = packageVersion(name_species) if (isProvided(params$custom_gmt)){ - database_string <- paste0("Also, a gprofiler query with this custom GMT file was done: `", basename(params$custom_gmt), "`. Entries of the following datasources were considered: ") - dbs <- paste(datasources, collapse="`, `") - database_string <- paste0(database_string, "`", dbs, "`.") -} else { + database_string <- paste0("Also, a gprofiler query with this custom GMT file was done: `", basename(params$custom_gmt), "`.") + + if (isProvided(params$datasources)) { + database_string <- paste(database_string, "Entries of the following datasources were considered: ") + dbs <- paste(datasources, collapse="`, `") + database_string <- paste0(database_string, "`", dbs, "`.") + } else { + database_string <- "" + } + +} else if (isProvided(params$datasources)) { database_string = "Also, the following databases were queried: " database_version_info <- get_version_info(params$organism) database_string <- paste0(database_string, "`gprofiler: ", database_version_info$gprofiler_version, "`") for (db in datasources) { database_string <- paste0(database_string, ", `", db, ": ", database_version_info$sources[[db]]$version, "`") } +} else { + database_string <- paste(database_string, "All datasources available in g:Profiler were considered (check https://biit.cs.ut.ee/gprofiler/gost and https://biit.cs.ut.ee/gprofiler/page/docs for more info).") } cat(paste0( diff --git a/bin/Execute_report.R b/bin/Execute_report.R index 5b9160d1..cd95b52e 100755 --- a/bin/Execute_report.R +++ b/bin/Execute_report.R @@ -34,6 +34,7 @@ option_list = list( make_option("--set_background", action="store_true", default=TRUE, help="Whether to use a background list for pathway analysis; if true, will only consider expressed genes (i.e. mean counts > 0) for PA."), make_option("--custom_background", type="character", default=NULL, help="Path to a custom background list TXT for pathway analysis; if provided, will only consider genes in that list for PA."), make_option(c("-w", "--min_DEG_pathway"), type="integer", default=NULL, help="min. number of genes DE in a pathway for this pathway to be considered enriched.", metavar="integer"), + make_option("--datasources", type="character", default=NULL, help="Which datasources to use for pathway analysis.", metavar="character"), make_option(c("-s", "--proj_summary"), type="character", default=NULL, help="Project summary file", metavar="character"), make_option(c("--path_quote"), type="character", default=NULL, help="Path to the quote PDF", metavar="character"), @@ -82,6 +83,7 @@ rmarkdown::render(opt$report, output_file = opt$output, knit_root_dir = wd, outp custom_background = opt$custom_background, keytype = opt$keytype, min_DEG_pathway = opt$min_DEG_pathway, + datasources = opt$datasources, path_proj_summary = opt$proj_summary, path_quote = opt$path_quote, diff --git a/conf/test.config b/conf/test.config index 05a27ad8..1b5562b4 100755 --- a/conf/test.config +++ b/conf/test.config @@ -30,4 +30,5 @@ params { run_pathway_analysis = true genome = 'GRCm38' quote = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/offer_example.pdf' + datasources = 'KEGG,REAC' } diff --git a/modules/local/report.nf b/modules/local/report.nf index eac509d0..4c5c9782 100755 --- a/modules/local/report.nf +++ b/modules/local/report.nf @@ -37,6 +37,7 @@ process REPORT { def custom_gmt_opt = custom_gmt.name != 'NO_FILE3' ? "--custom_gmt $custom_gmt" : '' def set_background_opt = params.set_background ? "--set_background TRUE" : "--set_background FALSE" def custom_background_opt = custom_background.name != 'NO_FILE7' ? "--custom_background $custom_background" : '' + def datasources_opt = params.datasources ? "--datasources $params.datasources" : '' def quote_opt = params.quote != 'NO_FILE5' ? "--path_quote $params.quote" : '' def software_versions_opt = params.software_versions != 'NO_FILE6' ? "--software_versions $params.software_versions" : '' @@ -76,6 +77,7 @@ process REPORT { --species_library $params.species_library \ --keytype $params.keytype \ --min_DEG_pathway $params.min_DEG_pathway \ + $datasources_opt \ $quote_opt \ $software_versions_opt \ --proj_summary $proj_summary \ diff --git a/nextflow.config b/nextflow.config index 9018f037..3838aeba 100755 --- a/nextflow.config +++ b/nextflow.config @@ -39,6 +39,7 @@ params { custom_gmt = 'NO_FILE3' set_background = true custom_background = 'NO_FILE7' + datasources = null // Additional args for the report project_summary = null diff --git a/nextflow_schema.json b/nextflow_schema.json index adddbf59..d41f8c26 100755 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -125,7 +125,7 @@ }, "custom_gmt": { "type": "string", - "description": "Path to custom GMT file for gost query, this allows e.g. to query older versions of databases for pathway analysis, not necessary if --skip_pathway_analysis = true." + "description": "Path to custom GMT file for gost query, this allows e.g. to query older versions of databases for pathway analysis, not necessary if --skip_pathway_analysis = true. If --datasources is set, the GMT file will be filtered for these datasources." }, "set_background": { "type": "boolean", @@ -140,6 +140,10 @@ "type": "integer", "default": 1, "description": "Integer indicating how many genes in a pathway must be differentially expressed to be considered as enriched, and report these pathways in tables and the final report. The default value is 1." + }, + "datasources": { + "type": "string", + "description": "Which datasources to use for pathway analysis, comma-separated string like 'KEGG,REAC'. See param 'sources' on https://rdrr.io/cran/gprofiler2/man/gost.html for a list of available sources. If not set, will use all sources. If set while a --custom_gmt is provided, will filter the GMT for these datasources (will not filter for the GO subtypes like GO:BP, just for GO)." } } }, From c77819213fc1e50b630a88950b71041573f0a48d Mon Sep 17 00:00:00 2001 From: WackerO Date: Tue, 20 Feb 2024 08:54:11 +0100 Subject: [PATCH 13/73] Updated changelog and test profiles --- CHANGELOG.md | 4 +++- assets/RNAseq_report.Rmd | 7 ++++++- conf/test.config | 1 - conf/test_custom_gmt.config | 1 + 4 files changed, 10 insertions(+), 3 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 7ada1e03..a9e2e139 100755 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -7,6 +7,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### Added +- [#225](https://github.com/qbic-pipelines/rnadeseq/pull/225) Added param for pathway analysis datasources - [#215](https://github.com/qbic-pipelines/rnadeseq/pull/215) Added gene_name to boxplot titles and filenames; increased threshold before overlapping PCA labels are hidden - [#213](https://github.com/qbic-pipelines/rnadeseq/pull/213) Added smrnaseq input support - [#212](https://github.com/qbic-pipelines/rnadeseq/pull/212) Added computational methods if no --software_versions @@ -17,6 +18,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### Changed +- [#225](https://github.com/qbic-pipelines/rnadeseq/pull/225) Shortened names of some output files - [#219](https://github.com/qbic-pipelines/rnadeseq/pull/219) Release 2.2 to master - [#220](https://github.com/qbic-pipelines/rnadeseq/pull/220) Commit suggestions from PR review for release 2.2 - [#218](https://github.com/qbic-pipelines/rnadeseq/pull/218) Preparing release 2.2 with version bumps @@ -34,7 +36,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### Fixed -- [#222](https://github.com/qbic-pipelines/rnadeseq/pull/222) Fixed too many devices error from tryCatch around normalized heatmaps +- [#225](https://github.com/qbic-pipelines/rnadeseq/pull/225) Fixed too many devices error from tryCatch around normalized heatmaps - [#212](https://github.com/qbic-pipelines/rnadeseq/pull/212) Fixed movability of interactive gostplots - [#208](https://github.com/qbic-pipelines/rnadeseq/pull/208) Fixed relevel bug, the function should now finally work! - [#207](https://github.com/qbic-pipelines/rnadeseq/pull/207) Fixed check of samples in counts vs metadata diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index 87f09691..d113be70 100755 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -1862,6 +1862,11 @@ if (mv_status) { } } +# Set datasources to NULL instead of c() to ensure that the gost queries work +if (!isProvided(params$datasources)) { + datasources <- NULL +} + # ------------------ # Set default params # ------------------ @@ -2474,7 +2479,7 @@ if (isProvided(params$custom_gmt)){ database_string <- paste0(database_string, ", `", db, ": ", database_version_info$sources[[db]]$version, "`") } } else { - database_string <- paste(database_string, "All datasources available in g:Profiler were considered (check https://biit.cs.ut.ee/gprofiler/gost and https://biit.cs.ut.ee/gprofiler/page/docs for more info).") + database_string <- paste("All datasources available in g:Profiler were considered (check https://biit.cs.ut.ee/gprofiler/gost and https://biit.cs.ut.ee/gprofiler/page/docs for more info).") } cat(paste0( diff --git a/conf/test.config b/conf/test.config index 1b5562b4..05a27ad8 100755 --- a/conf/test.config +++ b/conf/test.config @@ -30,5 +30,4 @@ params { run_pathway_analysis = true genome = 'GRCm38' quote = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/offer_example.pdf' - datasources = 'KEGG,REAC' } diff --git a/conf/test_custom_gmt.config b/conf/test_custom_gmt.config index e3f84934..345c2bef 100755 --- a/conf/test_custom_gmt.config +++ b/conf/test_custom_gmt.config @@ -30,4 +30,5 @@ params { run_pathway_analysis = true genome = 'GRCm38' custom_gmt = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/gprofiler_full_mmusculus.ENSG.gmt' + datasources = 'KEGG,REAC' } From d177582340bd128c6172c200655671af24dc7293 Mon Sep 17 00:00:00 2001 From: WackerO Date: Tue, 20 Feb 2024 10:35:10 +0100 Subject: [PATCH 14/73] bugfix, docu update --- assets/RNAseq_report.Rmd | 8 ++++---- docs/output.md | 2 +- 2 files changed, 5 insertions(+), 5 deletions(-) diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index d113be70..13aeb2ec 100755 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -2027,9 +2027,9 @@ for (file in contrast_files){ scale_fill_continuous(high = "#132B43", low = "#56B1F7") + ggtitle("Enriched pathways") + xlab("") + ylab("Gene fraction (DE genes / Pathway size)") - ggsave(p, filename = paste0("pathway_analysis", "/", fname, "/", db_source, "_pathway_enrichment_plot.pdf"), device = "pdf", height = 5+0.5*nrow(df_subset), units = "cm", limitsize=F) - ggsave(p, filename = paste0("pathway_analysis", "/", fname, "/", db_source, "_pathway_enrichment_plot.png"), device = "png", height = 5+0.5*nrow(df_subset), units = "cm", dpi = 300, limitsize=F) - ggsave(p, filename = paste0("pathway_analysis", "/", fname, "/", db_source, "_pathway_enrichment_plot.svg"), device = "svg", height = 5+0.5*nrow(df_subset), units = "cm", dpi = 300, limitsize=F) + ggsave(p, filename = paste0("pathway_analysis", "/", fname, "/", make.names(db_source), "_pathway_enrichment_plot.pdf"), device = "pdf", height = 5+0.5*nrow(df_subset), units = "cm", limitsize=F) + ggsave(p, filename = paste0("pathway_analysis", "/", fname, "/", make.names(db_source), "_pathway_enrichment_plot.png"), device = "png", height = 5+0.5*nrow(df_subset), units = "cm", dpi = 300, limitsize=F) + ggsave(p, filename = paste0("pathway_analysis", "/", fname, "/", make.names(db_source), "_pathway_enrichment_plot.svg"), device = "svg", height = 5+0.5*nrow(df_subset), units = "cm", dpi = 300, limitsize=F) # Plotting heatmaps and KEGG pathways for all pathways print("Plotting heatmaps...") @@ -2198,7 +2198,7 @@ Inside the pathway analysis results folder, a subfolder for each contrast used f Expand/collapse - `*_gost_pathway_venn_diagram.pdf/png` - Venn diagrams showing the numbers of enriched pathways when using a background gene list vs when not using a bg list. -- `*_KEGG_pathway_enrichment_plot.pdf/png` +- `*_pathway_enrichment_plot.pdf/png` - Barplots showing the proportion of differentially expressed genes in the pathway. - `KEGG_pathways/` - Contains the KEGG pathways graphs with the log fold change of the differentially expressed genes. diff --git a/docs/output.md b/docs/output.md index 99131726..47213b9e 100755 --- a/docs/output.md +++ b/docs/output.md @@ -52,7 +52,7 @@ If pathway analysis was run, this directory contains the zipped pathway analysis - `*_gost_pathway_venn_diagram.pdf/png` - Venn diagrams showing the numbers of enriched pathways when using a background gene list vs when not using a bg list. -- `*_KEGG_pathway_enrichment_plot.pdf/png` +- `*_pathway_enrichment_plot.pdf/png` - Barplots showing the proportion of differentially expressed genes in the pathway. - `KEGG_pathways/` - Contains the KEGG pathways graphs with the log fold change of the differentially expressed genes. From 803d56a1ab9d8b1de65bb05bf4c92adac2783633 Mon Sep 17 00:00:00 2001 From: WackerO Date: Tue, 20 Feb 2024 11:03:10 +0100 Subject: [PATCH 15/73] bugfix --- assets/RNAseq_report.Rmd | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index 13aeb2ec..eee83bd5 100755 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -2015,7 +2015,7 @@ for (file in contrast_files){ # Plotting results for df df_subset <- data.frame(Pathway_name = df$short_name, Pathway_code = df$term_id, DE_genes = df$intersection_size, Pathway_size = df$term_size, Fraction_DE = (df$intersection_size / df$term_size), Padj = df$p_value, DE_genes_names = df$intersection) write.table(df_subset, - file = paste0("pathway_analysis", "/", fname, "/", db_source, "_pathway_enrichment_results.tsv"), + file = paste0("pathway_analysis", "/", fname, "/", make.names(db_source), "_pathway_enrichment_results.tsv"), sep="\t", quote = F, col.names = T, row.names = F) # Enriched pathways horizontal barplots of padj values From 6b3199cfd6515d6b35d3f347275b4757b5004266 Mon Sep 17 00:00:00 2001 From: WackerO Date: Tue, 20 Feb 2024 13:22:08 +0100 Subject: [PATCH 16/73] file path changes --- assets/RNAseq_report.Rmd | 37 ++++++++++++++++++++----------------- docs/output.md | 6 ++++-- 2 files changed, 24 insertions(+), 19 deletions(-) diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index eee83bd5..daed4082 100755 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -1882,11 +1882,12 @@ theme_set(theme_classic()) for (file in contrast_files){ - #Reading DE genes list + # Reading DE genes list fname <- tools::file_path_sans_ext(basename(file)) dir.create(paste("pathway_analysis", fname, sep="/")) dir.create(paste("pathway_analysis", fname, "KEGG_pathways", sep="/")) + dir.create(paste("pathway_analysis", fname, "enrichment_plots", sep="/")) DE_genes <- read.csv(file = paste0(path_contrasts, file), sep="\t", header = T) DE_genes <- as.data.frame(DE_genes) @@ -1961,13 +1962,13 @@ for (file in contrast_files){ # Generate non-interactive pathway dotplots in the folder pg <- gostplot(gostres, capped=T, interactive=F) - ggsave(pg, filename = "pathway_analysis/gost_pathway_enrichment_plot.pdf", + ggsave(pg, filename = paste0("pathway_analysis/", "/", fname, "/gost_pathway_gostplot.pdf"), device="pdf", height=10, width=15, units="cm", limitsize=F) - ggsave(pg, filename = "pathway_analysis/gost_pathway_enrichment_plot.png", + ggsave(pg, filename = paste0("pathway_analysis/", "/", fname, "/gost_pathway_gostplot.png"), device="png", height=10, width=15, units="cm", dpi=300, limitsize=F) - ggsave(pg, filename = "pathway_analysis/gost_pathway_enrichment_plot.svg", + ggsave(pg, filename = paste0("pathway_analysis/", "/", fname, "/gost_pathway_gostplot.svg"), device="svg", height=10, width=15, units="cm", dpi=300, limitsize=F) @@ -1993,7 +1994,7 @@ for (file in contrast_files){ # Save pathway enrichment table in tsv format write.table(pathway_gostres_table, - file = paste0("pathway_analysis", "/", fname, "/", "pathway_enrichment_results.tsv"), + file = paste0("pathway_analysis", "/", fname, "/pathway_enrichment_results.tsv"), sep="\t", quote = F, col.names = T, row.names = F) print("------------------------------------") @@ -2027,9 +2028,9 @@ for (file in contrast_files){ scale_fill_continuous(high = "#132B43", low = "#56B1F7") + ggtitle("Enriched pathways") + xlab("") + ylab("Gene fraction (DE genes / Pathway size)") - ggsave(p, filename = paste0("pathway_analysis", "/", fname, "/", make.names(db_source), "_pathway_enrichment_plot.pdf"), device = "pdf", height = 5+0.5*nrow(df_subset), units = "cm", limitsize=F) - ggsave(p, filename = paste0("pathway_analysis", "/", fname, "/", make.names(db_source), "_pathway_enrichment_plot.png"), device = "png", height = 5+0.5*nrow(df_subset), units = "cm", dpi = 300, limitsize=F) - ggsave(p, filename = paste0("pathway_analysis", "/", fname, "/", make.names(db_source), "_pathway_enrichment_plot.svg"), device = "svg", height = 5+0.5*nrow(df_subset), units = "cm", dpi = 300, limitsize=F) + ggsave(p, filename = paste0("pathway_analysis", "/", fname, "/enrichment_plots/", make.names(db_source), "_pathway_enrichment_plot.pdf"), device = "pdf", height = 5+0.5*nrow(df_subset), units = "cm", limitsize=F) + ggsave(p, filename = paste0("pathway_analysis", "/", fname, "/enrichment_plots/", make.names(db_source), "_pathway_enrichment_plot.png"), device = "png", height = 5+0.5*nrow(df_subset), units = "cm", dpi = 300, limitsize=F) + ggsave(p, filename = paste0("pathway_analysis", "/", fname, "/enrichment_plots/", make.names(db_source), "_pathway_enrichment_plot.svg"), device = "svg", height = 5+0.5*nrow(df_subset), units = "cm", dpi = 300, limitsize=F) # Plotting heatmaps and KEGG pathways for all pathways print("Plotting heatmaps...") @@ -2054,11 +2055,11 @@ for (file in contrast_files){ # Because the image devices sometimes don't work here, surround them with a tryCatch so that the code continues even if a certain heatmap is not saved tryCatch( { - png(filename = paste0("pathway_analysis", "/",fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, ".png"), width = ifelse(nrow(mat)<=20, 200, 125)+ncol(mat)*40, height = 50+nrow(mat)*50) + png(filename = paste0("pathway_analysis", "/",fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", make.names(pathway$term_id), ".png"), width = ifelse(nrow(mat)<=20, 200, 125)+ncol(mat)*40, height = 50+nrow(mat)*50) pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T) }, error=function(e) { - print(paste0("Could not save pathway_analysis", "/",fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, ".png because of the following error:\n", e)) + print(paste0("Could not save pathway_analysis", "/",fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", make.names(pathway$term_id), ".png because of the following error:\n", e)) }, finally={ dev.off() @@ -2066,11 +2067,11 @@ for (file in contrast_files){ ) tryCatch( { - pdf(paste0("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, ".pdf"), width = ifelse(nrow(mat)<=20, 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005) + pdf(paste0("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", make.names(pathway$term_id), ".pdf"), width = ifelse(nrow(mat)<=20, 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005) pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T) }, error=function(e) { - print(paste0("Could not save pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, ".pdf because of the following error:\n", e)) + print(paste0("Could not save pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", make.names(pathway$term_id), ".pdf because of the following error:\n", e)) }, finally={ dev.off() @@ -2078,11 +2079,11 @@ for (file in contrast_files){ ) tryCatch( { - svg(paste0("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, ".svg"), width = ifelse(nrow(mat)<=20, 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005) + svg(paste0("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", make.names(pathway$term_id), ".svg"), width = ifelse(nrow(mat)<=20, 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005) pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T) }, error=function(e) { - print(paste0("Could not save pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, ".svg because of the following error:\n", e)) + print(paste0("Could not save pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", make.names(pathway$term_id), ".svg because of the following error:\n", e)) }, finally={ dev.off() @@ -2197,9 +2198,11 @@ Inside the pathway analysis results folder, a subfolder for each contrast used f

Expand/collapse - `*_gost_pathway_venn_diagram.pdf/png` - - Venn diagrams showing the numbers of enriched pathways when using a background gene list vs when not using a bg list. -- `*_pathway_enrichment_plot.pdf/png` - - Barplots showing the proportion of differentially expressed genes in the pathway. + - Venn diagrams showing the numbers of enriched pathways when using a background gene list vs when not using a bg list. +- `enrichment_plots/*_pathway_enrichment_plot.{pdf/png/svg}` + - Barplots showing the proportion of differentially expressed genes in the pathway for a certain pathway database. +- `gost_pathway_gostplot.{pdf/png/svg}` + - Manhattan plots displaying all enriched pathways. - `KEGG_pathways/` - Contains the KEGG pathways graphs with the log fold change of the differentially expressed genes. - `pathway_heatmaps` diff --git a/docs/output.md b/docs/output.md index 47213b9e..6cc4e044 100755 --- a/docs/output.md +++ b/docs/output.md @@ -52,8 +52,10 @@ If pathway analysis was run, this directory contains the zipped pathway analysis - `*_gost_pathway_venn_diagram.pdf/png` - Venn diagrams showing the numbers of enriched pathways when using a background gene list vs when not using a bg list. -- `*_pathway_enrichment_plot.pdf/png` - - Barplots showing the proportion of differentially expressed genes in the pathway. +- `enrichment_plots/*_pathway_enrichment_plot.{pdf/png/svg}` + - Barplots showing the proportion of differentially expressed genes in the pathway for a certain pathway database. +- `gost_pathway_gostplot.{pdf/png/svg}` + - Manhattan plots displaying all enriched pathways. - `KEGG_pathways/` - Contains the KEGG pathways graphs with the log fold change of the differentially expressed genes. - `pathway_heatmaps` From 469724bde42129c728ecd0a2a50e6a22cbc11b97 Mon Sep 17 00:00:00 2001 From: WackerO Date: Tue, 20 Feb 2024 14:22:37 +0100 Subject: [PATCH 17/73] updated test ymls --- tests/test.yml | 32 ++++++++++++++++---------------- tests/test_custom_gmt.yml | 4 ++-- tests/test_full.yml | 22 +++++++++++----------- tests/test_smrnaseq.yml | 20 ++++++++++---------- 4 files changed, 39 insertions(+), 39 deletions(-) diff --git a/tests/test.yml b/tests/test.yml index 56ff217d..bb988d70 100755 --- a/tests/test.yml +++ b/tests/test.yml @@ -60,35 +60,35 @@ - path: results_test/differential_gene_expression/plots/PCA_plot.pdf - path: results_test/differential_gene_expression/plots/PCA_plot.png - path: results_test/differential_gene_expression/plots/PCA_plot.svg - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_venn_diagram.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_venn_diagram.png - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_venn_diagram.svg - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_pathway_enrichment_results.tsv + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_venn_diagram.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_venn_diagram.png + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_venn_diagram.svg + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_enrichment_plot.png + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO/pathway_enrichment_results.tsv md5sum: be0f292a205f2ecd453ee27ecdc62615 - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_results.tsv + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathway_enrichment_results.tsv md5sum: f0e6d8117e3ff05ddabc3abb98f514b7 - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_results.tsv + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO/REAC_pathway_enrichment_results.tsv md5sum: 3d7616fd0f6cb3aaf9f08252b2bcd223 - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_plot.png + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/enrichment_plots/KEGG_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/enrichment_plots/KEGG_pathway_enrichment_plot.png + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/enrichment_plots/REAC_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/enrichment_plots/REAC_pathway_enrichment_plot.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04360.DE_contrast_condition_genotype_WT_vs_KO.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04360.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04360.xml - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04610.DE_contrast_condition_genotype_WT_vs_KO.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04610.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04610.xml - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_pathway_enrichment_results.tsv + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/pathway_enrichment_results.tsv md5sum: 0bd6dfd32bad6f5292f6cde91f019844 - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_results.tsv + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/REAC_pathway_enrichment_results.tsv md5sum: 452ca5dd8d5f190648988502f85d3ef1 - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.pdf - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.png - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.svg - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_plot.png + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/REAC_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/REAC_pathway_enrichment_plot.png - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf - path: results_test/RNAseq_report.html diff --git a/tests/test_custom_gmt.yml b/tests/test_custom_gmt.yml index 589ccf13..6baa345d 100755 --- a/tests/test_custom_gmt.yml +++ b/tests/test_custom_gmt.yml @@ -62,9 +62,9 @@ - path: results_test/differential_gene_expression/plots/PCA_plot.svg - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.pdf - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_pathway_enrichment_results.tsv + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/pathway_enrichment_results.tsv md5sum: eae154d9c7a526cc8c3fcd1a586fa0ed - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_pathway_enrichment_results.tsv + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/pathway_enrichment_results.tsv md5sum: bd2cbbf85c3e241743e60b9b8263a31c - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf - path: results_test/RNAseq_report.html diff --git a/tests/test_full.yml b/tests/test_full.yml index c4477cbf..c6aa2bc8 100755 --- a/tests/test_full.yml +++ b/tests/test_full.yml @@ -62,24 +62,24 @@ - path: results_test/differential_gene_expression/plots/PCA_plot.svg - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.pdf - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_pathway_enrichment_results.tsv + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/pathway_enrichment_results.tsv md5sum: be0f292a205f2ecd453ee27ecdc62615 - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_results.tsv + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathway_enrichment_results.tsv md5sum: f0e6d8117e3ff05ddabc3abb98f514b7 - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_results.tsv + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/REAC_pathway_enrichment_results.tsv md5sum: 3d7616fd0f6cb3aaf9f08252b2bcd223 - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_plot.png + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathway_enrichment_plot.png + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/REAC_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/REAC_pathway_enrichment_plot.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04360.DE_contrast_condition_genotype_WT_vs_KO.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04360.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04360.xml - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_pathway_enrichment_results.tsv + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/pathway_enrichment_results.tsv md5sum: 0bd6dfd32bad6f5292f6cde91f019844 - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_results.tsv + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/REAC_pathway_enrichment_results.tsv md5sum: 452ca5dd8d5f190648988502f85d3ef1 - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_plot.png + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/REAC_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/REAC_pathway_enrichment_plot.png - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf - path: results_test/RNAseq_report.html diff --git a/tests/test_smrnaseq.yml b/tests/test_smrnaseq.yml index 34ff104b..3b430223 100755 --- a/tests/test_smrnaseq.yml +++ b/tests/test_smrnaseq.yml @@ -72,18 +72,18 @@ - path: results_test/differential_gene_expression/plots/PCA_plot_with_labels.pdf - path: results_test/differential_gene_expression/plots/PCA_plot_with_labels.png - path: results_test/differential_gene_expression/plots/PCA_plot_with_labels.svg - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/DE_contrast_condition_treatment_clone9_vs_clone1_KEGG_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/DE_contrast_condition_treatment_clone9_vs_clone1_KEGG_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/DE_contrast_condition_treatment_clone9_vs_clone1_KEGG_pathway_enrichment_plot.svg - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/DE_contrast_condition_treatment_clone9_vs_clone1_KEGG_pathway_enrichment_results.tsv + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/enrichment_plots/KEGG_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/enrichment_plots/KEGG_pathway_enrichment_plot.png + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/enrichment_plots/KEGG_pathway_enrichment_plot.svg + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/KEGG_pathway_enrichment_results.tsv md5sum: 2637f5f6f4e74e7723280449e3ac647b - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/DE_contrast_condition_treatment_clone9_vs_clone1_pathway_enrichment_results.tsv + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/pathway_enrichment_results.tsv md5sum: a091e2ae9a678599d9e994082907d646 - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/DE_contrast_condition_treatment_control_vs_clone1_KEGG_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/DE_contrast_condition_treatment_control_vs_clone1_KEGG_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/DE_contrast_condition_treatment_control_vs_clone1_KEGG_pathway_enrichment_plot.svg - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/DE_contrast_condition_treatment_control_vs_clone1_KEGG_pathway_enrichment_results.tsv + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/KEGG_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/KEGG_pathway_enrichment_plot.png + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/KEGG_pathway_enrichment_plot.svg + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/KEGG_pathway_enrichment_results.tsv md5sum: 497cbe026880c9ea78c5591c7c4d07f7 - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/DE_contrast_condition_treatment_control_vs_clone1_pathway_enrichment_results.tsv + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/pathway_enrichment_results.tsv md5sum: 63a2adf7c4a971dc7443503cf968e455 - path: results_test/RNAseq_report.html From 4a472f845d165f2e66fa18d179c9590baac59c9b Mon Sep 17 00:00:00 2001 From: WackerO Date: Tue, 20 Feb 2024 16:09:10 +0100 Subject: [PATCH 18/73] more yml fixing, pinned validation plugin version --- assets/RNAseq_report.Rmd | 28 ++++++++++++++-------------- nextflow.config | 5 +++++ tests/test.yml | 15 ++++++++------- tests/test_custom_gmt.yml | 8 ++++++-- tests/test_full.yml | 18 +++++++++--------- tests/test_smrnaseq.yml | 14 +++++++------- tests/test_star_salmon.yml | 4 ++-- 7 files changed, 51 insertions(+), 41 deletions(-) diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index daed4082..87b7928f 100755 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -169,7 +169,7 @@ isProvided <- function(value) { #this function is used to produce a more informative error message when count table and metadata table disagree write_error_msg <- function(counts, qbicCodes) { error_msg <- "Count table headers do not exactly match the metadata table sample names!\n" - + counts_msg <- "" counts_offending <- "" for (c in names(counts)) { @@ -178,7 +178,7 @@ write_error_msg <- function(counts, qbicCodes) { counts_offending <- paste0(counts_offending, c, "\n") } } - + meta_msg <- "" meta_offending <- "" for (m in qbicCodes) { @@ -399,7 +399,7 @@ if (params$input_type == "smrnaseq") { missing_files <- paste0(substring(missing_files, 1, nchar(missing_files)-2), ".") stop(missing_files, call.=F) } - + # Combine files for later files <- c(hairpin_files, other_files) @@ -1054,7 +1054,7 @@ cat(paste0("*** The raw count table and normalized count tables are available [here](./differential_gene_expression/gene_counts_tables). The differential expression analysis is performed using the raw gene count table. -For PCA analysis and heatmap plotting, the `r norm_method_long_text` normalized gene counts were used. +For PCA analysis and heatmap plotting, the `r norm_method_long_text` normalized gene counts were used. `r norm_method_override_text` `r nsub_genes_text` @@ -1653,7 +1653,7 @@ if (isProvided(params$path_genelist)) { ggsave(filename=paste("differential_gene_expression/plots/boxplots_requested_genes/",requested_genes_plot_gene_name[i],"_",requested_genes_plot_Ensembl[i],".svg",sep=""), width=10, height=5, plot=plot) ggsave(filename=paste("differential_gene_expression/plots/boxplots_requested_genes/",requested_genes_plot_gene_name[i],"_",requested_genes_plot_Ensembl[i],".png",sep=""), width=10, height=5, plot=plot) ggsave(filename=paste("differential_gene_expression/plots/boxplots_requested_genes/",requested_genes_plot_gene_name[i],"_",requested_genes_plot_Ensembl[i],".pdf",sep=""), width=10, height=5, plot=plot) - } + } } ``` @@ -1720,7 +1720,7 @@ if (isProvided(params$path_genelist)) { ggsave(filename=paste0("differential_gene_expression/plots/boxplots_requested_genes/",requested_genes_plot_gene_name[i],"_",requested_genes_plot_Ensembl[i],"_after_batchcorrect.svg"), width=10, height=5, plot=plot) ggsave(filename=paste0("differential_gene_expression/plots/boxplots_requested_genes/",requested_genes_plot_gene_name[i],"_",requested_genes_plot_Ensembl[i],"_after_batchcorrect.png"), width=10, height=5, plot=plot) ggsave(filename=paste0("differential_gene_expression/plots/boxplots_requested_genes/",requested_genes_plot_gene_name[i],"_",requested_genes_plot_Ensembl[i],"_after_batchcorrect.pdf"), width=10, height=5, plot=plot) - } + } } ``` @@ -1912,11 +1912,11 @@ for (file in contrast_files){ custom_bg=custom_background, domain_scope="custom_annotated" ) - + pathway_gostres <- gostres$result pathway_gostres <- as.data.frame(pathway_gostres[which(pathway_gostres$significant==TRUE),]) pathway_gostres <- pathway_gostres[which(pathway_gostres$intersection_size>=min_DEG_pathway),] - + # Also run without bg for Venn diagrams gostres_nobg <- gost(query=q, organism=gost_id, @@ -1982,9 +1982,9 @@ for (file in contrast_files){ no_background=pathway_gostres_nobg$term_name ) pv <- ggvenn::ggvenn(VennInput) - ggsave(paste0("pathway_analysis", "/", fname, "_gost_pathway_venn_diagram.pdf"),pv, device="pdf", width=10, height=15) - ggsave(paste0("pathway_analysis", "/", fname, "_gost_pathway_venn_diagram.png"),pv, device="png", width=10, height=15) - ggsave(paste0("pathway_analysis", "/", fname, "_gost_pathway_venn_diagram.svg"),pv, device="svg", width=10, height=15) + ggsave(paste0("pathway_analysis", "/", fname, "/gost_pathway_venn_diagram.pdf"),pv, device="pdf", width=10, height=15) + ggsave(paste0("pathway_analysis", "/", fname, "/gost_pathway_venn_diagram.png"),pv, device="png", width=10, height=15) + ggsave(paste0("pathway_analysis", "/", fname, "/gost_pathway_venn_diagram.svg"),pv, device="svg", width=10, height=15) } } @@ -2052,7 +2052,7 @@ for (file in contrast_files){ if (!isProvided(params$citest) & nrow(mat)>1){ dir.create(paste("pathway_analysis", fname, "pathway_heatmaps", sep="/")) - # Because the image devices sometimes don't work here, surround them with a tryCatch so that the code continues even if a certain heatmap is not saved + # Because the image devices sometimes don't work here, surround them with a tryCatch so that the code continues even if a certain heatmap is not saved tryCatch( { png(filename = paste0("pathway_analysis", "/",fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", make.names(pathway$term_id), ".png"), width = ifelse(nrow(mat)<=20, 200, 125)+ncol(mat)*40, height = 50+nrow(mat)*50) @@ -2241,7 +2241,7 @@ names(q_list) <- q_names if (length(q_list) > 0) { #gost query - + if (isProvided(params$set_background)) { gostres <- gost(query=q_list, organism=gost_id, @@ -2452,7 +2452,7 @@ as.character(params$revision), "` pipeline [^5], which was written using the nf-core template [@ewels2020nf]. For differential expression analysis, the read quantification data resulting from `", quant_tool, "` were processed with the R package `DESeq2 v", packageVersion("DESeq2"), -"` [@love2014differential]. The thresholds for differentially expressed genes were set to ", +"` [@love2014differential]. The thresholds for differentially expressed genes were set to ", pval_text, " for the p-value and ", logFC_text, " for the log2 Fold Change." )) diff --git a/nextflow.config b/nextflow.config index 3838aeba..afce37a5 100755 --- a/nextflow.config +++ b/nextflow.config @@ -6,6 +6,11 @@ ---------------------------------------------------------------------------------------- */ +// Pin nf-validation to prevent errors from new versions +plugins { + id 'nf-validation@1.1.3' +} + // Global default params, used in configs // params marked with //* exist for the sake of completeness only; // enabling them prevents igenomes from working diff --git a/tests/test.yml b/tests/test.yml index bb988d70..c08f7e0a 100755 --- a/tests/test.yml +++ b/tests/test.yml @@ -63,13 +63,14 @@ - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_venn_diagram.pdf - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_venn_diagram.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_venn_diagram.svg - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO/pathway_enrichment_results.tsv + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_gostplot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_gostplot.png + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_gostplot.svg + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/pathway_enrichment_results.tsv md5sum: be0f292a205f2ecd453ee27ecdc62615 - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathway_enrichment_results.tsv + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathway_enrichment_results.tsv md5sum: f0e6d8117e3ff05ddabc3abb98f514b7 - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO/REAC_pathway_enrichment_results.tsv + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/REAC_pathway_enrichment_results.tsv md5sum: 3d7616fd0f6cb3aaf9f08252b2bcd223 - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/enrichment_plots/KEGG_pathway_enrichment_plot.pdf - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/enrichment_plots/KEGG_pathway_enrichment_plot.png @@ -88,7 +89,7 @@ - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.pdf - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.png - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.svg - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/REAC_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/REAC_pathway_enrichment_plot.png + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/enrichment_plots/REAC_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/enrichment_plots/REAC_pathway_enrichment_plot.png - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf - path: results_test/RNAseq_report.html diff --git a/tests/test_custom_gmt.yml b/tests/test_custom_gmt.yml index 6baa345d..ea161a15 100755 --- a/tests/test_custom_gmt.yml +++ b/tests/test_custom_gmt.yml @@ -60,8 +60,12 @@ - path: results_test/differential_gene_expression/plots/PCA_plot.pdf - path: results_test/differential_gene_expression/plots/PCA_plot.png - path: results_test/differential_gene_expression/plots/PCA_plot.svg - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.png + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_gostplot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_gostplot.png + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_gostplot.svg + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_venn_diagram.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_venn_diagram.png + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_venn_diagram.svg - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/pathway_enrichment_results.tsv md5sum: eae154d9c7a526cc8c3fcd1a586fa0ed - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/pathway_enrichment_results.tsv diff --git a/tests/test_full.yml b/tests/test_full.yml index c6aa2bc8..9ec30e7f 100755 --- a/tests/test_full.yml +++ b/tests/test_full.yml @@ -60,18 +60,18 @@ - path: results_test/differential_gene_expression/plots/PCA_plot.pdf - path: results_test/differential_gene_expression/plots/PCA_plot.png - path: results_test/differential_gene_expression/plots/PCA_plot.svg - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.png + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_gostplot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_gostplot.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/pathway_enrichment_results.tsv - md5sum: be0f292a205f2ecd453ee27ecdc62615 + md5sum: 21b907d52008f66f9a85190400fd211e - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathway_enrichment_results.tsv - md5sum: f0e6d8117e3ff05ddabc3abb98f514b7 + md5sum: 92115e662e0e2489ad05d28f3981fa3a - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/REAC_pathway_enrichment_results.tsv - md5sum: 3d7616fd0f6cb3aaf9f08252b2bcd223 - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/REAC_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/REAC_pathway_enrichment_plot.png + md5sum: 9077c63139668fd01d3f4086e6acb7bc + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/enrichment_plots/KEGG_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/enrichment_plots/KEGG_pathway_enrichment_plot.png + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/enrichment_plots/REAC_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/enrichment_plots/REAC_pathway_enrichment_plot.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04360.DE_contrast_condition_genotype_WT_vs_KO.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04360.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04360.xml diff --git a/tests/test_smrnaseq.yml b/tests/test_smrnaseq.yml index 3b430223..39676c01 100755 --- a/tests/test_smrnaseq.yml +++ b/tests/test_smrnaseq.yml @@ -76,14 +76,14 @@ - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/enrichment_plots/KEGG_pathway_enrichment_plot.png - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/enrichment_plots/KEGG_pathway_enrichment_plot.svg - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/KEGG_pathway_enrichment_results.tsv - md5sum: 2637f5f6f4e74e7723280449e3ac647b + md5sum: 326f3495ca42dbc527484cef3ee0e58d - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/pathway_enrichment_results.tsv - md5sum: a091e2ae9a678599d9e994082907d646 - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/KEGG_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/KEGG_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/KEGG_pathway_enrichment_plot.svg + md5sum: b77e2839aa95ce3a9562d03a13629eb4 + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/enrichment_plots/KEGG_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/enrichment_plots/KEGG_pathway_enrichment_plot.png + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/enrichment_plots/KEGG_pathway_enrichment_plot.svg - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/KEGG_pathway_enrichment_results.tsv - md5sum: 497cbe026880c9ea78c5591c7c4d07f7 + md5sum: 64b3a3716b779900e99f56688c4b27e8 - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/pathway_enrichment_results.tsv - md5sum: 63a2adf7c4a971dc7443503cf968e455 + md5sum: 05e83209e36face6ead8b066155fdbe5 - path: results_test/RNAseq_report.html diff --git a/tests/test_star_salmon.yml b/tests/test_star_salmon.yml index dbe49c9a..f657920b 100755 --- a/tests/test_star_salmon.yml +++ b/tests/test_star_salmon.yml @@ -69,8 +69,8 @@ - path: results_test/differential_gene_expression/plots/PCA_plot.pdf - path: results_test/differential_gene_expression/plots/PCA_plot.png - path: results_test/differential_gene_expression/plots/PCA_plot.svg - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_pathway_enrichment_results.tsv + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/pathway_enrichment_results.tsv md5sum: cd967cc355045d9dc8c0bb6ffcf40b61 - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_pathway_enrichment_results.tsv + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/pathway_enrichment_results.tsv md5sum: 68b329da9893e34099c7d8ad5cb9c940 - path: results_test/RNAseq_report.html From 6869d813d32f8cbd4f85a2899c3b087554045f95 Mon Sep 17 00:00:00 2001 From: WackerO Date: Wed, 21 Feb 2024 14:52:07 +0100 Subject: [PATCH 19/73] more yml updates, trying to get rsem to run --- assets/RNAseq_report.Rmd | 32 ++++++++++++++++++++++++++++++++ tests/test.yml | 16 ++++++++-------- tests/test_full.yml | 8 ++++---- tests/test_star_salmon.yml | 4 ++-- 4 files changed, 46 insertions(+), 14 deletions(-) diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index 87b7928f..c30ae187 100755 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -529,6 +529,7 @@ DT::datatable(df_QA_neat, options = list(scrollX = "100%", scrollY = "400px", p if (params$input_type %in% c("featurecounts", "smrnaseq")) { cds <- DESeqDataSetFromMatrix( countData =count.table, colData =metadata, design = eval(parse(text=as.character(design[[1]])))) } else if (params$input_type %in% c("rsem", "salmon")) { + write("rsem1", file="/home-link/iivow01/git/rnadeseq/error/status", append=F) ## Create a dataframe which consists of both the gene id and the transcript name gtf <- rtracklayer::import(params$path_gtf) gtf <- as.data.frame(gtf, header=T) @@ -573,6 +574,7 @@ if (params$input_type %in% c("featurecounts", "smrnaseq")) { coldata <- data.frame(files = files, names= qbicCodes, sampleconditions) coldata$combfactor <- metadata$combfactor rownames(coldata) <- NULL + write("rsem2", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) #Do tximeta, this is necessary to run DESeq on rsem (imports the rsem output #and modifies them as they are not integers and can not directly be used for cds) @@ -585,6 +587,7 @@ if (params$input_type %in% c("featurecounts", "smrnaseq")) { #dds from SummarizedExperiment , then run DESeq cds <- DESeqDataSet(se, design = as.formula(eval(parse(text=as.character(design[[1]]))))) + write("rsem3", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) # Load salmon count files } else if (params$input_type == "salmon") { @@ -625,12 +628,16 @@ if (params$input_type %in% c("featurecounts", "smrnaseq")) { #Apply relevel if provided; this must be done before calling DESeq() if (isProvided(params$path_relevel)) { + write("rsem4", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) + relevel_table <- read.table(params$path_relevel, sep="\t", header = T, colClasses = "character") write.table(relevel_table, file="differential_gene_expression/metadata/relevel.tsv") for (i in c(1:nrow(relevel_table))) { relev <- relevel_table[i,] cds[[paste(relev[1])]] <- relevel(cds[[paste(relev[1])]], paste(relev[2])) } + write("rsem5", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) + } # Run actual DESeq analysis @@ -644,6 +651,8 @@ if (params$input_type %in% c("featurecounts", "smrnaseq")) { nm <- assays(cds)[["avgTxLength"]] sf <- estimateSizeFactorsForMatrix(counts(cds)/nm) } + write("rsem6", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) + write.table(sf,paste("differential_gene_expression/gene_counts_tables/sizeFactor_libraries.tsv",sep=""), append = FALSE, quote = FALSE, sep = "\t",eol = "\n", na = "NA", dec = ".", row.names = T, col.names = F, qmethod = c("escape", "double")) # Write cds assay table to file @@ -716,6 +725,7 @@ if (run_rlog){ rownames(norm_counts) <- norm_counts$Ensembl_ID } } + write("rsem7", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) ############################## DIAGNOSTICS AND QUALITY CONTROL PLOTS ############################### # Cooks distances: get important for example when checking knock-out and overexpression studies @@ -771,6 +781,7 @@ par(mfrow = c(1, 3)) meanSdPlot(log2(counts(cds,normalized=TRUE)[notAllZero,] + 1),ylab = "sd raw count data") meanSdPlot(assay((if (run_rlog) rld else vsd)[notAllZero,]),ylab = paste("sd", norm_method_text, "transformed count data")) dev.off() + write("rsem8", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) # Further diagnostics plots res=0 @@ -859,6 +870,8 @@ for (i in resultsNames(cds)[-1]) { dev.off() rm(res,qs,bins,ratios,use,h1,h2,colori) } + write("rsem9", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) + ``` @@ -951,6 +964,8 @@ The read mapping statistics to the reference genome for each sample are shown be # Disable auto-conversion of paths to relative paths as we provide include_graphics() with absolute paths. Also, error=F is necessary in every include_graphic() call to not fail with an error knitr.graphics.rel_path = F knitr::include_graphics(paste0(wd, "/QC/multiqc_plots/svg/mqc_star_alignment_plot_1_pc.svg"), error=F) + write("rsem10", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) + ``` ```{r STAR_readnums,echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/"))} @@ -1102,6 +1117,8 @@ if (run_rlog) { } else { vsd_noncorrected <- vsd } + write("rsem11", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) + ``` \ @@ -1315,6 +1332,7 @@ if (isProvided(params$path_contrast_list)) { } } write.table(contrasts, file="differential_gene_expression/metadata/contrast_list.tsv", quote=F) + write("rsem12", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) ## Contrast calculation for contrast list for (i in c(1:nrow(contrasts))) { @@ -1433,6 +1451,7 @@ if (!isProvided(params$path_contrast_matrix) & !isProvided(params$path_contrast_ DE_genes_df = cbind(DE_genes_df,results_DEseq_contrast) } } + write("rsem13", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) # Write contrast names to file write(contrast_names, file="contrast_names.txt", sep="\t") @@ -1559,6 +1578,7 @@ DE_all$logpval[DE_all$logpval > 16] <- 17 DE_all <- na.omit(DE_all) log2FoldChange_min <- min(DE_all$log2FoldChange) log2FoldChange_max <- max(DE_all$log2FoldChange) + write("rsem14", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) # get quotient of log2FoldChange range and 10 to determine if xticks of plot need to be scaled discretely (otherwise, for large FC ranges, there will be too many xticks and they will overlap) log2FoldChange_quotient <- length(seq(log2FoldChange_min, log2FoldChange_max, 10)) @@ -1691,6 +1711,7 @@ for (i in nrow(random_DE_genes_plot)) { ggsave(filename=paste0("differential_gene_expression/plots/boxplots_example_genes/",genename,"_",ensid,"_after_batchcorrect.png"), width=10, height=5, plot=plot) ggsave(filename=paste0("differential_gene_expression/plots/boxplots_example_genes/",genename,"_",ensid,"_after_batchcorrect.pdf"), width=10, height=5, plot=plot) } + write("rsem15", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) # make boxplots of interesting genes in gene list if (isProvided(params$path_genelist)) { @@ -1780,6 +1801,7 @@ if (isProvided(params$datasources)) { } else { datasources <- c() } + write("rsem16", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) # Default to 1 for the nb of genes DE in a pathway min_DEG_pathway <- as.integer(params$min_DEG_pathway) @@ -1851,6 +1873,7 @@ if (isProvided(params$custom_gmt)){ } ) } +write("rsem17", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) # If GMT exist(s), create metadata folder and move GMT(s) there @@ -1861,6 +1884,7 @@ if (mv_status) { system(paste("cp", params$custom_gmt, "pathway_analysis/metadata/")) } } + write("rsem17.1", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) # Set datasources to NULL instead of c() to ensure that the gost queries work if (!isProvided(params$datasources)) { @@ -1879,6 +1903,7 @@ theme_set(theme_classic()) # ---------------------- # For each contrast do pathway analysis + write("rsem17.2", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) for (file in contrast_files){ @@ -1890,6 +1915,7 @@ for (file in contrast_files){ dir.create(paste("pathway_analysis", fname, "enrichment_plots", sep="/")) DE_genes <- read.csv(file = paste0(path_contrasts, file), sep="\t", header = T) DE_genes <- as.data.frame(DE_genes) + write("rsem17.3", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) # Skip pathway analysis for the contrast if not 2 or more DE genes were found if (nrow(DE_genes) < 2){ @@ -1899,6 +1925,7 @@ for (file in contrast_files){ # Define list of Ensemble IDs (q) to run the pathway analysis q = as.character(DE_genes$Ensembl_ID) + write("rsem17.4", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) # gost query if (isProvided(params$set_background)) { @@ -1916,6 +1943,7 @@ for (file in contrast_files){ pathway_gostres <- gostres$result pathway_gostres <- as.data.frame(pathway_gostres[which(pathway_gostres$significant==TRUE),]) pathway_gostres <- pathway_gostres[which(pathway_gostres$intersection_size>=min_DEG_pathway),] + write("rsem17.5", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) # Also run without bg for Venn diagrams gostres_nobg <- gost(query=q, @@ -1930,6 +1958,7 @@ for (file in contrast_files){ pathway_gostres_nobg <- gostres_nobg$result pathway_gostres_nobg <- as.data.frame(pathway_gostres_nobg[which(pathway_gostres_nobg$significant==TRUE),]) pathway_gostres_nobg <- pathway_gostres_nobg[which(pathway_gostres_nobg$intersection_size>=min_DEG_pathway),] + write("rsem17.6", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) } else { gostres <- gost(query=q, @@ -1943,6 +1972,7 @@ for (file in contrast_files){ ) } + write("rsem18", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) # Make data frame of gost result pathway_gostres <- gostres$result @@ -2160,6 +2190,7 @@ if (isProvided(params$path_genelist)){ mat$gene_name <- NULL mat$Ensembl_ID <- NULL mat <- data.matrix(mat) + write("rsem19", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) if (nrow(mat)>1){ png(filename = paste0("pathway_analysis", "/", genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.png"), width = 100+ncol(mat)*50, height = 50+nrow(mat)*100) @@ -2383,6 +2414,7 @@ cat("No software versions file was provided for primary analysis.") ``` ```{r software_versions_RNASeq, echo=FALSE, results='asis', eval=isProvided(params$path_software_versions)} + write("rsem20", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) # In the following part, I use newlines (in code) where possible whenever the paste0 function switches from a string to a variable call to better distinguish between the two so it is clearer in which lines quotes are needed and in which not --> Cannot always use newlines as this sometimes translates into the report.html diff --git a/tests/test.yml b/tests/test.yml index c08f7e0a..585f4a04 100755 --- a/tests/test.yml +++ b/tests/test.yml @@ -67,11 +67,11 @@ - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_gostplot.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_gostplot.svg - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/pathway_enrichment_results.tsv - md5sum: be0f292a205f2ecd453ee27ecdc62615 + md5sum: 21b907d52008f66f9a85190400fd211e - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathway_enrichment_results.tsv - md5sum: f0e6d8117e3ff05ddabc3abb98f514b7 + md5sum: 92115e662e0e2489ad05d28f3981fa3a - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/REAC_pathway_enrichment_results.tsv - md5sum: 3d7616fd0f6cb3aaf9f08252b2bcd223 + md5sum: 9077c63139668fd01d3f4086e6acb7bc - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/enrichment_plots/KEGG_pathway_enrichment_plot.pdf - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/enrichment_plots/KEGG_pathway_enrichment_plot.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/enrichment_plots/REAC_pathway_enrichment_plot.pdf @@ -83,12 +83,12 @@ - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04610.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04610.xml - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/pathway_enrichment_results.tsv - md5sum: 0bd6dfd32bad6f5292f6cde91f019844 + md5sum: bdc0167b8b33a0da44e34768eead8376 - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/REAC_pathway_enrichment_results.tsv - md5sum: 452ca5dd8d5f190648988502f85d3ef1 - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.png - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.svg + md5sum: 6594fdb8c0eb591fa131982a8be00877 + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/gost_pathway_venn_diagram.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/gost_pathway_venn_diagram.png + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/gost_pathway_venn_diagram.svg - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/enrichment_plots/REAC_pathway_enrichment_plot.pdf - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/enrichment_plots/REAC_pathway_enrichment_plot.png - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf diff --git a/tests/test_full.yml b/tests/test_full.yml index 9ec30e7f..beed5c7d 100755 --- a/tests/test_full.yml +++ b/tests/test_full.yml @@ -76,10 +76,10 @@ - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04360.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04360.xml - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/pathway_enrichment_results.tsv - md5sum: 0bd6dfd32bad6f5292f6cde91f019844 + md5sum: bdc0167b8b33a0da44e34768eead8376 - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/REAC_pathway_enrichment_results.tsv - md5sum: 452ca5dd8d5f190648988502f85d3ef1 - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/REAC_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/REAC_pathway_enrichment_plot.png + md5sum: 6594fdb8c0eb591fa131982a8be00877 + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/enrichment_plots/REAC_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/enrichment_plots/REAC_pathway_enrichment_plot.png - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf - path: results_test/RNAseq_report.html diff --git a/tests/test_star_salmon.yml b/tests/test_star_salmon.yml index f657920b..9d693acb 100755 --- a/tests/test_star_salmon.yml +++ b/tests/test_star_salmon.yml @@ -70,7 +70,7 @@ - path: results_test/differential_gene_expression/plots/PCA_plot.png - path: results_test/differential_gene_expression/plots/PCA_plot.svg - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/pathway_enrichment_results.tsv - md5sum: cd967cc355045d9dc8c0bb6ffcf40b61 + md5sum: 30af65ef53ef7e389e116c31d31bd1d6 - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/pathway_enrichment_results.tsv - md5sum: 68b329da9893e34099c7d8ad5cb9c940 + md5sum: 4a3e61c2b4fea94c327acc63c83c0a4a - path: results_test/RNAseq_report.html From 9655686c56d3056abf7a81b4c6f81d26b6f48fca Mon Sep 17 00:00:00 2001 From: WackerO Date: Thu, 22 Feb 2024 09:22:24 +0100 Subject: [PATCH 20/73] Fixed a bug with finally dev.off; restricted rsem pathway analysis to GO:CC to speed it up --- assets/RNAseq_report.Rmd | 13 ++++--------- conf/test_star_rsem.config | 1 + 2 files changed, 5 insertions(+), 9 deletions(-) diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index c30ae187..a6920e4f 100755 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -2087,38 +2087,33 @@ for (file in contrast_files){ { png(filename = paste0("pathway_analysis", "/",fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", make.names(pathway$term_id), ".png"), width = ifelse(nrow(mat)<=20, 200, 125)+ncol(mat)*40, height = 50+nrow(mat)*50) pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T) + dev.off() }, error=function(e) { print(paste0("Could not save pathway_analysis", "/",fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", make.names(pathway$term_id), ".png because of the following error:\n", e)) - }, - finally={ - dev.off() } ) tryCatch( { pdf(paste0("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", make.names(pathway$term_id), ".pdf"), width = ifelse(nrow(mat)<=20, 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005) pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T) + dev.off() }, error=function(e) { print(paste0("Could not save pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", make.names(pathway$term_id), ".pdf because of the following error:\n", e)) - }, - finally={ - dev.off() } ) tryCatch( { svg(paste0("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", make.names(pathway$term_id), ".svg"), width = ifelse(nrow(mat)<=20, 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005) pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T) + dev.off() }, error=function(e) { print(paste0("Could not save pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", make.names(pathway$term_id), ".svg because of the following error:\n", e)) - }, - finally={ - dev.off() } ) + graphics.off() # Close all still open devices from errors. Tried doing this in finally, but that doesn't work } # Plotting pathway view only for kegg pathways diff --git a/conf/test_star_rsem.config b/conf/test_star_rsem.config index 03c11cd5..5e633e22 100755 --- a/conf/test_star_rsem.config +++ b/conf/test_star_rsem.config @@ -27,6 +27,7 @@ params { input_type = 'rsem' run_pathway_analysis = true genome = 'GRCh37' + datasources = 'GO:CC' // species_library = "org.Hs.eg.db" // organism = "hsapiens" // keytype = "ENSEMBL" From cf4a195e162d9311a9df8a34f8cf22105ded6ba3 Mon Sep 17 00:00:00 2001 From: WackerO Date: Thu, 22 Feb 2024 10:50:51 +0100 Subject: [PATCH 21/73] removed local writes --- assets/RNAseq_report.Rmd | 33 +-------------------------------- 1 file changed, 1 insertion(+), 32 deletions(-) diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index a6920e4f..ae01a018 100755 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -529,7 +529,7 @@ DT::datatable(df_QA_neat, options = list(scrollX = "100%", scrollY = "400px", p if (params$input_type %in% c("featurecounts", "smrnaseq")) { cds <- DESeqDataSetFromMatrix( countData =count.table, colData =metadata, design = eval(parse(text=as.character(design[[1]])))) } else if (params$input_type %in% c("rsem", "salmon")) { - write("rsem1", file="/home-link/iivow01/git/rnadeseq/error/status", append=F) + ## Create a dataframe which consists of both the gene id and the transcript name gtf <- rtracklayer::import(params$path_gtf) gtf <- as.data.frame(gtf, header=T) @@ -574,7 +574,6 @@ if (params$input_type %in% c("featurecounts", "smrnaseq")) { coldata <- data.frame(files = files, names= qbicCodes, sampleconditions) coldata$combfactor <- metadata$combfactor rownames(coldata) <- NULL - write("rsem2", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) #Do tximeta, this is necessary to run DESeq on rsem (imports the rsem output #and modifies them as they are not integers and can not directly be used for cds) @@ -587,7 +586,6 @@ if (params$input_type %in% c("featurecounts", "smrnaseq")) { #dds from SummarizedExperiment , then run DESeq cds <- DESeqDataSet(se, design = as.formula(eval(parse(text=as.character(design[[1]]))))) - write("rsem3", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) # Load salmon count files } else if (params$input_type == "salmon") { @@ -628,16 +626,12 @@ if (params$input_type %in% c("featurecounts", "smrnaseq")) { #Apply relevel if provided; this must be done before calling DESeq() if (isProvided(params$path_relevel)) { - write("rsem4", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) - relevel_table <- read.table(params$path_relevel, sep="\t", header = T, colClasses = "character") write.table(relevel_table, file="differential_gene_expression/metadata/relevel.tsv") for (i in c(1:nrow(relevel_table))) { relev <- relevel_table[i,] cds[[paste(relev[1])]] <- relevel(cds[[paste(relev[1])]], paste(relev[2])) } - write("rsem5", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) - } # Run actual DESeq analysis @@ -651,7 +645,6 @@ if (params$input_type %in% c("featurecounts", "smrnaseq")) { nm <- assays(cds)[["avgTxLength"]] sf <- estimateSizeFactorsForMatrix(counts(cds)/nm) } - write("rsem6", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) write.table(sf,paste("differential_gene_expression/gene_counts_tables/sizeFactor_libraries.tsv",sep=""), append = FALSE, quote = FALSE, sep = "\t",eol = "\n", na = "NA", dec = ".", row.names = T, col.names = F, qmethod = c("escape", "double")) @@ -725,7 +718,6 @@ if (run_rlog){ rownames(norm_counts) <- norm_counts$Ensembl_ID } } - write("rsem7", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) ############################## DIAGNOSTICS AND QUALITY CONTROL PLOTS ############################### # Cooks distances: get important for example when checking knock-out and overexpression studies @@ -781,7 +773,6 @@ par(mfrow = c(1, 3)) meanSdPlot(log2(counts(cds,normalized=TRUE)[notAllZero,] + 1),ylab = "sd raw count data") meanSdPlot(assay((if (run_rlog) rld else vsd)[notAllZero,]),ylab = paste("sd", norm_method_text, "transformed count data")) dev.off() - write("rsem8", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) # Further diagnostics plots res=0 @@ -870,8 +861,6 @@ for (i in resultsNames(cds)[-1]) { dev.off() rm(res,qs,bins,ratios,use,h1,h2,colori) } - write("rsem9", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) - ``` @@ -964,8 +953,6 @@ The read mapping statistics to the reference genome for each sample are shown be # Disable auto-conversion of paths to relative paths as we provide include_graphics() with absolute paths. Also, error=F is necessary in every include_graphic() call to not fail with an error knitr.graphics.rel_path = F knitr::include_graphics(paste0(wd, "/QC/multiqc_plots/svg/mqc_star_alignment_plot_1_pc.svg"), error=F) - write("rsem10", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) - ``` ```{r STAR_readnums,echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/"))} @@ -1117,8 +1104,6 @@ if (run_rlog) { } else { vsd_noncorrected <- vsd } - write("rsem11", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) - ``` \ @@ -1332,7 +1317,6 @@ if (isProvided(params$path_contrast_list)) { } } write.table(contrasts, file="differential_gene_expression/metadata/contrast_list.tsv", quote=F) - write("rsem12", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) ## Contrast calculation for contrast list for (i in c(1:nrow(contrasts))) { @@ -1451,7 +1435,6 @@ if (!isProvided(params$path_contrast_matrix) & !isProvided(params$path_contrast_ DE_genes_df = cbind(DE_genes_df,results_DEseq_contrast) } } - write("rsem13", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) # Write contrast names to file write(contrast_names, file="contrast_names.txt", sep="\t") @@ -1578,7 +1561,6 @@ DE_all$logpval[DE_all$logpval > 16] <- 17 DE_all <- na.omit(DE_all) log2FoldChange_min <- min(DE_all$log2FoldChange) log2FoldChange_max <- max(DE_all$log2FoldChange) - write("rsem14", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) # get quotient of log2FoldChange range and 10 to determine if xticks of plot need to be scaled discretely (otherwise, for large FC ranges, there will be too many xticks and they will overlap) log2FoldChange_quotient <- length(seq(log2FoldChange_min, log2FoldChange_max, 10)) @@ -1711,7 +1693,6 @@ for (i in nrow(random_DE_genes_plot)) { ggsave(filename=paste0("differential_gene_expression/plots/boxplots_example_genes/",genename,"_",ensid,"_after_batchcorrect.png"), width=10, height=5, plot=plot) ggsave(filename=paste0("differential_gene_expression/plots/boxplots_example_genes/",genename,"_",ensid,"_after_batchcorrect.pdf"), width=10, height=5, plot=plot) } - write("rsem15", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) # make boxplots of interesting genes in gene list if (isProvided(params$path_genelist)) { @@ -1801,7 +1782,6 @@ if (isProvided(params$datasources)) { } else { datasources <- c() } - write("rsem16", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) # Default to 1 for the nb of genes DE in a pathway min_DEG_pathway <- as.integer(params$min_DEG_pathway) @@ -1873,7 +1853,6 @@ if (isProvided(params$custom_gmt)){ } ) } -write("rsem17", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) # If GMT exist(s), create metadata folder and move GMT(s) there @@ -1884,7 +1863,6 @@ if (mv_status) { system(paste("cp", params$custom_gmt, "pathway_analysis/metadata/")) } } - write("rsem17.1", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) # Set datasources to NULL instead of c() to ensure that the gost queries work if (!isProvided(params$datasources)) { @@ -1903,7 +1881,6 @@ theme_set(theme_classic()) # ---------------------- # For each contrast do pathway analysis - write("rsem17.2", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) for (file in contrast_files){ @@ -1915,7 +1892,6 @@ for (file in contrast_files){ dir.create(paste("pathway_analysis", fname, "enrichment_plots", sep="/")) DE_genes <- read.csv(file = paste0(path_contrasts, file), sep="\t", header = T) DE_genes <- as.data.frame(DE_genes) - write("rsem17.3", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) # Skip pathway analysis for the contrast if not 2 or more DE genes were found if (nrow(DE_genes) < 2){ @@ -1925,7 +1901,6 @@ for (file in contrast_files){ # Define list of Ensemble IDs (q) to run the pathway analysis q = as.character(DE_genes$Ensembl_ID) - write("rsem17.4", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) # gost query if (isProvided(params$set_background)) { @@ -1943,7 +1918,6 @@ for (file in contrast_files){ pathway_gostres <- gostres$result pathway_gostres <- as.data.frame(pathway_gostres[which(pathway_gostres$significant==TRUE),]) pathway_gostres <- pathway_gostres[which(pathway_gostres$intersection_size>=min_DEG_pathway),] - write("rsem17.5", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) # Also run without bg for Venn diagrams gostres_nobg <- gost(query=q, @@ -1958,7 +1932,6 @@ for (file in contrast_files){ pathway_gostres_nobg <- gostres_nobg$result pathway_gostres_nobg <- as.data.frame(pathway_gostres_nobg[which(pathway_gostres_nobg$significant==TRUE),]) pathway_gostres_nobg <- pathway_gostres_nobg[which(pathway_gostres_nobg$intersection_size>=min_DEG_pathway),] - write("rsem17.6", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) } else { gostres <- gost(query=q, @@ -1972,8 +1945,6 @@ for (file in contrast_files){ ) } - write("rsem18", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) - # Make data frame of gost result pathway_gostres <- gostres$result @@ -2185,7 +2156,6 @@ if (isProvided(params$path_genelist)){ mat$gene_name <- NULL mat$Ensembl_ID <- NULL mat <- data.matrix(mat) - write("rsem19", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) if (nrow(mat)>1){ png(filename = paste0("pathway_analysis", "/", genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.png"), width = 100+ncol(mat)*50, height = 50+nrow(mat)*100) @@ -2409,7 +2379,6 @@ cat("No software versions file was provided for primary analysis.") ``` ```{r software_versions_RNASeq, echo=FALSE, results='asis', eval=isProvided(params$path_software_versions)} - write("rsem20", file="/home-link/iivow01/git/rnadeseq/error/status", append=T) # In the following part, I use newlines (in code) where possible whenever the paste0 function switches from a string to a variable call to better distinguish between the two so it is clearer in which lines quotes are needed and in which not --> Cannot always use newlines as this sometimes translates into the report.html From 8df1b3186d7763319e4be31958b1cfcb9ed494fb Mon Sep 17 00:00:00 2001 From: WackerO Date: Tue, 27 Feb 2024 14:10:27 +0100 Subject: [PATCH 22/73] Fixed rsem test --- tests/test_star_rsem.yml | 70 ++++++++++++++++++++++------------------ 1 file changed, 38 insertions(+), 32 deletions(-) diff --git a/tests/test_star_rsem.yml b/tests/test_star_rsem.yml index c9cbdd49..6566048b 100755 --- a/tests/test_star_rsem.yml +++ b/tests/test_star_rsem.yml @@ -75,38 +75,44 @@ - path: results_test/differential_gene_expression/plots/PCA_plot.pdf - path: results_test/differential_gene_expression/plots/PCA_plot.png - path: results_test/differential_gene_expression/plots/PCA_plot.svg - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_KEGG_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_KEGG_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_KEGG_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_KEGG_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_KEGG_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_KEGG_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_REAC_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_REAC_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_REAC_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_REAC_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_REAC_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_REAC_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_pathway_enrichment_results.tsv + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.png + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.svg + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/gost_pathway_gostplot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/gost_pathway_gostplot.png + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/gost_pathway_gostplot.svg + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/gost_pathway_venn_diagram.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/gost_pathway_venn_diagram.png + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/gost_pathway_venn_diagram.svg + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/pathway_enrichment_results.tsv md5sum: b1c0a1374c963fff71083a9cdefe8e9b - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_pathway_enrichment_results.tsv + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/GO.CC_pathway_enrichment_results.tsv + md5sum: 2bbbf2de30f9da36e4c5b2df7a4ef5e7 + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.png + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.svg + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/gost_pathway_gostplot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/gost_pathway_gostplot.png + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/gost_pathway_gostplot.svg + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/gost_pathway_venn_diagram.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/gost_pathway_venn_diagram.png + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/gost_pathway_venn_diagram.svg + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/pathway_enrichment_results.tsv md5sum: 631e89b4de9042936fa2652bde036e7d - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_pathway_enrichment_results.tsv + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/GO.CC_pathway_enrichment_results.tsv + md5sum: 48a7c37c08b19fe71cfee34ae7c3fd8b + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.png + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.svg + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/gost_pathway_gostplot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/gost_pathway_gostplot.png + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/gost_pathway_gostplot.svg + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/gost_pathway_venn_diagram.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/gost_pathway_venn_diagram.png + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/gost_pathway_venn_diagram.svg + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/pathway_enrichment_results.tsv md5sum: 80123c4ba2c97eaa0217816f41122855 - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_KEGG_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_KEGG_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_KEGG_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_KEGG_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_KEGG_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_KEGG_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_REAC_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_REAC_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_REAC_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_REAC_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_REAC_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_REAC_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_KEGG_pathway_enrichment_results.tsv - md5sum: 276e871c81e85217bbd7eef951676084 - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_REAC_pathway_enrichment_results.tsv - md5sum: 9e6847fe5d631f09305dfa00dd3472d1 - - path: results_test/RNAseq_report.html + - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf + - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.png + - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.svg + - path: results_test/pathway_analysis/metadata/gprofiler_full_hsapiens.ENSG.gmt From db3111ccebdfc8434881728051b5021f5ff2f785 Mon Sep 17 00:00:00 2001 From: WackerO Date: Tue, 27 Feb 2024 15:08:01 +0100 Subject: [PATCH 23/73] md5sum update --- tests/test_star_rsem.yml | 9 ++++----- 1 file changed, 4 insertions(+), 5 deletions(-) diff --git a/tests/test_star_rsem.yml b/tests/test_star_rsem.yml index 6566048b..d0d4b11c 100755 --- a/tests/test_star_rsem.yml +++ b/tests/test_star_rsem.yml @@ -85,7 +85,7 @@ - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/gost_pathway_venn_diagram.png - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/gost_pathway_venn_diagram.svg - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/pathway_enrichment_results.tsv - md5sum: b1c0a1374c963fff71083a9cdefe8e9b + md5sum: 9eef4eab3da88b4b7e465f982be72acb - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/GO.CC_pathway_enrichment_results.tsv md5sum: 2bbbf2de30f9da36e4c5b2df7a4ef5e7 - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.pdf @@ -98,9 +98,9 @@ - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/gost_pathway_venn_diagram.png - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/gost_pathway_venn_diagram.svg - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/pathway_enrichment_results.tsv - md5sum: 631e89b4de9042936fa2652bde036e7d + md5sum: cbda28eeba947c53847ea28fdaa0cebd - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/GO.CC_pathway_enrichment_results.tsv - md5sum: 48a7c37c08b19fe71cfee34ae7c3fd8b + md5sum: 6b79ce64567f94ad3fc492e1a56fce58 - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.pdf - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.png - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.svg @@ -111,8 +111,7 @@ - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/gost_pathway_venn_diagram.png - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/gost_pathway_venn_diagram.svg - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/pathway_enrichment_results.tsv - md5sum: 80123c4ba2c97eaa0217816f41122855 + md5sum: 68f919b58d7fe5cc1b9a38665abccf78 - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf - - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.png - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.svg - path: results_test/pathway_analysis/metadata/gprofiler_full_hsapiens.ENSG.gmt From 69855ac7d082403d0719b2359e273710eff060d6 Mon Sep 17 00:00:00 2001 From: WackerO Date: Thu, 29 Feb 2024 11:12:18 +0100 Subject: [PATCH 24/73] changed versions to dev --- .github/workflows/ci.yml | 8 ++++---- Dockerfile | 6 +++--- environment.yml | 2 +- modules/local/report.nf | 2 +- nextflow.config | 2 +- 5 files changed, 10 insertions(+), 10 deletions(-) diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index e40f2eb6..ae4653ab 100755 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -40,14 +40,14 @@ jobs: environment.yml - name: Build new docker image if: env.MATCHED_FILES - run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:2.2 + run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:2.3dev # Change the version above and the third version below before/after release - name: Pull docker image if: ${{ !env.MATCHED_FILES }} run: | docker pull ghcr.io/qbic-pipelines/rnadeseq:dev - docker tag ghcr.io/qbic-pipelines/rnadeseq:dev ghcr.io/qbic-pipelines/rnadeseq:2.2 + docker tag ghcr.io/qbic-pipelines/rnadeseq:dev ghcr.io/qbic-pipelines/rnadeseq:2.3dev - name: Install Nextflow uses: nf-core/setup-nextflow@v1 @@ -94,14 +94,14 @@ jobs: environment.yml - name: Build new docker image if: env.MATCHED_FILES - run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:2.2 + run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:2.3dev # Change the version above and the third version below before/after release - name: Pull docker image if: ${{ !env.MATCHED_FILES }} run: | docker pull ghcr.io/qbic-pipelines/rnadeseq:dev - docker tag ghcr.io/qbic-pipelines/rnadeseq:dev ghcr.io/qbic-pipelines/rnadeseq:2.2 + docker tag ghcr.io/qbic-pipelines/rnadeseq:dev ghcr.io/qbic-pipelines/rnadeseq:2.3dev - name: Install Nextflow uses: nf-core/setup-nextflow@v1 diff --git a/Dockerfile b/Dockerfile index c13267b3..1a3b6a0f 100755 --- a/Dockerfile +++ b/Dockerfile @@ -5,14 +5,14 @@ LABEL org.opencontainers.image.authors="Gisela Gabernet, Alexander Peltzer, Oska LABEL org.opencontainers.image.licenses=MIT COPY environment.yml / #RUN conda install -c conda-forge mamba -RUN mamba env create --file /environment.yml -p /opt/conda/envs/qbic-pipelines-rnadeseq-2.2 && \ +RUN mamba env create --file /environment.yml -p /opt/conda/envs/qbic-pipelines-rnadeseq-2.3dev && \ mamba clean --all --yes RUN apt-get update -qq && \ apt-get install -y zip procps ghostscript # Add conda installation dir to PATH -ENV PATH /opt/conda/envs/qbic-pipelines-rnadeseq-2.2/bin:$PATH +ENV PATH /opt/conda/envs/qbic-pipelines-rnadeseq-2.3dev/bin:$PATH # Dump the details of the installed packates to a file for posterity -RUN mamba env export --name qbic-pipelines-rnadeseq-2.2 > qbic-pipelines-rnadeseq-2.2.yml +RUN mamba env export --name qbic-pipelines-rnadeseq-2.3dev > qbic-pipelines-rnadeseq-2.3dev.yml # Instruct R processes to use these empty files instead of clashing with a local config RUN touch .Rprofile RUN touch .Renviron diff --git a/environment.yml b/environment.yml index cf3dc64d..a9447dcb 100755 --- a/environment.yml +++ b/environment.yml @@ -1,7 +1,7 @@ # You can use this file to create a conda environment for this pipeline: # conda env create -f environment.yml # use this to find packages: https://anaconda.org/ -name: qbic-pipelines-rnadeseq-2.2 +name: qbic-pipelines-rnadeseq-2.3dev channels: - bioconda - conda-forge diff --git a/modules/local/report.nf b/modules/local/report.nf index 4c5c9782..bc145c70 100755 --- a/modules/local/report.nf +++ b/modules/local/report.nf @@ -1,6 +1,6 @@ process REPORT { - container 'ghcr.io/qbic-pipelines/rnadeseq:2.2' + container 'ghcr.io/qbic-pipelines/rnadeseq:2.3dev' input: path gene_counts diff --git a/nextflow.config b/nextflow.config index afce37a5..7397f9c5 100755 --- a/nextflow.config +++ b/nextflow.config @@ -258,7 +258,7 @@ manifest { description = """Differential gene expression analysis and pathway analysis of RNAseq data""" mainScript = 'main.nf' nextflowVersion = '!>=23.04.0' - version = '2.2' + version = '2.3dev' doi = '' } From 114ecfa656990e743b885c91388acd5c284a19fc Mon Sep 17 00:00:00 2001 From: qbicStefanC Date: Tue, 26 Mar 2024 09:29:38 +0100 Subject: [PATCH 25/73] Update RNAseq_report.Rmd Update RNAseq_report.Rmd for DE information --- assets/RNAseq_report.Rmd | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index b1aa99b8..7feb1f6c 100644 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -2423,7 +2423,7 @@ as.character(params$revision), quant_tool, "` were processed with the R package `DESeq2 v", packageVersion("DESeq2"), "` [@love2014differential]. The thresholds for differentially expressed genes were set to ", -pval_text, " for the p-value and ", +pval_text, " for the adjusted p-value and ", logFC_text, " for the log2 Fold Change." )) From 962c18530ce0831561bcfbb3bff4311a36e038c0 Mon Sep 17 00:00:00 2001 From: WackerO Date: Wed, 10 Apr 2024 15:55:06 +0200 Subject: [PATCH 26/73] fixed version of container, fixed venn path in fulltest --- .github/workflows/ci.yml | 8 ++++---- modules/local/report.nf | 2 +- tests/test_full.yml | 6 +++--- 3 files changed, 8 insertions(+), 8 deletions(-) diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index ae4653ab..d1e1401f 100755 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -40,14 +40,14 @@ jobs: environment.yml - name: Build new docker image if: env.MATCHED_FILES - run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:2.3dev + run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:dev # Change the version above and the third version below before/after release - name: Pull docker image if: ${{ !env.MATCHED_FILES }} run: | docker pull ghcr.io/qbic-pipelines/rnadeseq:dev - docker tag ghcr.io/qbic-pipelines/rnadeseq:dev ghcr.io/qbic-pipelines/rnadeseq:2.3dev + docker tag ghcr.io/qbic-pipelines/rnadeseq:dev ghcr.io/qbic-pipelines/rnadeseq:dev - name: Install Nextflow uses: nf-core/setup-nextflow@v1 @@ -94,14 +94,14 @@ jobs: environment.yml - name: Build new docker image if: env.MATCHED_FILES - run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:2.3dev + run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:dev # Change the version above and the third version below before/after release - name: Pull docker image if: ${{ !env.MATCHED_FILES }} run: | docker pull ghcr.io/qbic-pipelines/rnadeseq:dev - docker tag ghcr.io/qbic-pipelines/rnadeseq:dev ghcr.io/qbic-pipelines/rnadeseq:2.3dev + docker tag ghcr.io/qbic-pipelines/rnadeseq:dev ghcr.io/qbic-pipelines/rnadeseq:dev - name: Install Nextflow uses: nf-core/setup-nextflow@v1 diff --git a/modules/local/report.nf b/modules/local/report.nf index bc145c70..a05ef3b7 100755 --- a/modules/local/report.nf +++ b/modules/local/report.nf @@ -1,6 +1,6 @@ process REPORT { - container 'ghcr.io/qbic-pipelines/rnadeseq:2.3dev' + container 'ghcr.io/qbic-pipelines/rnadeseq:dev' input: path gene_counts diff --git a/tests/test_full.yml b/tests/test_full.yml index a59a3305..7b4759eb 100755 --- a/tests/test_full.yml +++ b/tests/test_full.yml @@ -82,7 +82,7 @@ - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/enrichment_plots/REAC_pathway_enrichment_plot.pdf - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/enrichment_plots/REAC_pathway_enrichment_plot.png - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.png - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.svg + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/gost_pathway_venn_diagram.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/gost_pathway_venn_diagram.png + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/gost_pathway_venn_diagram.svg - path: results_test/RNAseq_report.html From c5f09692e6f0b4924e908f7b72320263f372a692 Mon Sep 17 00:00:00 2001 From: WackerO Date: Thu, 11 Apr 2024 15:14:28 +0200 Subject: [PATCH 27/73] struggling with an error --- .github/workflows/ci.yml | 8 +- Dockerfile | 6 +- assets/RNAseq_report.Rmd | 315 ++++++++++++++++++++++++++++----------- environment.yml | 2 +- modules/local/report.nf | 2 +- 5 files changed, 235 insertions(+), 98 deletions(-) diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index e40f2eb6..d1e1401f 100644 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -40,14 +40,14 @@ jobs: environment.yml - name: Build new docker image if: env.MATCHED_FILES - run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:2.2 + run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:dev # Change the version above and the third version below before/after release - name: Pull docker image if: ${{ !env.MATCHED_FILES }} run: | docker pull ghcr.io/qbic-pipelines/rnadeseq:dev - docker tag ghcr.io/qbic-pipelines/rnadeseq:dev ghcr.io/qbic-pipelines/rnadeseq:2.2 + docker tag ghcr.io/qbic-pipelines/rnadeseq:dev ghcr.io/qbic-pipelines/rnadeseq:dev - name: Install Nextflow uses: nf-core/setup-nextflow@v1 @@ -94,14 +94,14 @@ jobs: environment.yml - name: Build new docker image if: env.MATCHED_FILES - run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:2.2 + run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:dev # Change the version above and the third version below before/after release - name: Pull docker image if: ${{ !env.MATCHED_FILES }} run: | docker pull ghcr.io/qbic-pipelines/rnadeseq:dev - docker tag ghcr.io/qbic-pipelines/rnadeseq:dev ghcr.io/qbic-pipelines/rnadeseq:2.2 + docker tag ghcr.io/qbic-pipelines/rnadeseq:dev ghcr.io/qbic-pipelines/rnadeseq:dev - name: Install Nextflow uses: nf-core/setup-nextflow@v1 diff --git a/Dockerfile b/Dockerfile index c13267b3..4f8ce7fe 100644 --- a/Dockerfile +++ b/Dockerfile @@ -5,14 +5,14 @@ LABEL org.opencontainers.image.authors="Gisela Gabernet, Alexander Peltzer, Oska LABEL org.opencontainers.image.licenses=MIT COPY environment.yml / #RUN conda install -c conda-forge mamba -RUN mamba env create --file /environment.yml -p /opt/conda/envs/qbic-pipelines-rnadeseq-2.2 && \ +RUN mamba env create --file /environment.yml -p /opt/conda/envs/qbic-pipelines-rnadeseq-dev && \ mamba clean --all --yes RUN apt-get update -qq && \ apt-get install -y zip procps ghostscript # Add conda installation dir to PATH -ENV PATH /opt/conda/envs/qbic-pipelines-rnadeseq-2.2/bin:$PATH +ENV PATH /opt/conda/envs/qbic-pipelines-rnadeseq-dev/bin:$PATH # Dump the details of the installed packates to a file for posterity -RUN mamba env export --name qbic-pipelines-rnadeseq-2.2 > qbic-pipelines-rnadeseq-2.2.yml +RUN mamba env export --name qbic-pipelines-rnadeseq-dev > qbic-pipelines-rnadeseq-dev.yml # Instruct R processes to use these empty files instead of clashing with a local config RUN touch .Rprofile RUN touch .Renviron diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index b1aa99b8..3cd6ad23 100644 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -43,6 +43,7 @@ params: custom_gmt: '' set_background: '' custom_background: '' + datasources: '' #Additional args for the report path_proj_summary: '' @@ -168,7 +169,7 @@ isProvided <- function(value) { #this function is used to produce a more informative error message when count table and metadata table disagree write_error_msg <- function(counts, qbicCodes) { error_msg <- "Count table headers do not exactly match the metadata table sample names!\n" - + counts_msg <- "" counts_offending <- "" for (c in names(counts)) { @@ -177,7 +178,7 @@ write_error_msg <- function(counts, qbicCodes) { counts_offending <- paste0(counts_offending, c, "\n") } } - + meta_msg <- "" meta_offending <- "" for (m in qbicCodes) { @@ -348,6 +349,7 @@ Auf der Morgenstelle 10, 72076 Tuebingen ```{r read_metadata, echo=FALSE, message=FALSE, warning=FALSE, results='asis'} +now <- Sys.time() # Load metadata: sample preparations tsv file from qPortal metadata <- read.table(params$path_metadata, sep="\t", header=TRUE,na.strings =c("","NaN"), quote=NULL, stringsAsFactors=F, dec=".", fill=TRUE, row.names=1) if (params$pathway_analysis) { @@ -398,7 +400,7 @@ if (params$input_type == "smrnaseq") { missing_files <- paste0(substring(missing_files, 1, nchar(missing_files)-2), ".") stop(missing_files, call.=F) } - + # Combine files for later files <- c(hairpin_files, other_files) @@ -494,6 +496,7 @@ if(file.exists(QAfile)){ } else { no_RIN <- TRUE } +write(paste((Sys.time() - now)/60, "min used for read_metadata"), file="/home-link/iivow01/git/rnadeseq/error/times", append=F) ``` ```{r RIN_block, echo=FALSE, results='asis', eval=RIN} @@ -522,12 +525,13 @@ DT::datatable(df_QA_neat, options = list(scrollX = "100%", scrollY = "400px", p ```{r DESeq, echo=FALSE, message=FALSE, warning=FALSE, results = 'hide'} - +now <- Sys.time() ################## RUN DESEQ2 ###################################### # Run DESeq function if (params$input_type %in% c("featurecounts", "smrnaseq")) { cds <- DESeqDataSetFromMatrix( countData =count.table, colData =metadata, design = eval(parse(text=as.character(design[[1]])))) } else if (params$input_type %in% c("rsem", "salmon")) { + ## Create a dataframe which consists of both the gene id and the transcript name gtf <- rtracklayer::import(params$path_gtf) gtf <- as.data.frame(gtf, header=T) @@ -599,7 +603,8 @@ if (params$input_type %in% c("featurecounts", "smrnaseq")) { } } missing_files <- paste0(substring(missing_files, 1, nchar(missing_files)-2), ".") - stop(missing_files, call.=F) } + stop(missing_files, call.=F) + } #The following steps are necessary for the processing of salmon output as the files do # not contain integer counts and can therefore not be directly used for cds @@ -634,6 +639,8 @@ if (isProvided(params$path_relevel)) { # Run actual DESeq analysis cds <- DESeq(cds) +write(paste((Sys.time() - now)/60, "min used for run DESeq2"), file="/home-link/iivow01/git/rnadeseq/error/times", append=T) +now <- Sys.time() # sizeFactors(cds) as indicator of library sequencing depth if (params$input_type %in% c("featurecounts", "smrnaseq")) { sf <- sizeFactors(cds) @@ -642,6 +649,7 @@ if (params$input_type %in% c("featurecounts", "smrnaseq")) { nm <- assays(cds)[["avgTxLength"]] sf <- estimateSizeFactorsForMatrix(counts(cds)/nm) } + write.table(sf,paste("differential_gene_expression/gene_counts_tables/sizeFactor_libraries.tsv",sep=""), append = FALSE, quote = FALSE, sep = "\t",eol = "\n", na = "NA", dec = ".", row.names = T, col.names = F, qmethod = c("escape", "double")) # Write cds assay table to file @@ -691,35 +699,43 @@ norm_method_text <- ifelse(run_rlog, "rlog", "vst") if (run_rlog){ # rlog transformation + write.table(assays(cds)[["counts"]], file="/home-link/iivow01/git/rnadeseq/error/counts_before_rlog.tsv", sep="\t", quote=F) rld <- rlog(cds, blind=FALSE) rld_names <- merge(x=gene_names, y=assay(rld), by.x = "Ensembl_ID", by.y="row.names") write.table(rld_names, "differential_gene_expression/gene_counts_tables/rlog_transformed_gene_counts.tsv", append = FALSE, quote = FALSE, sep = "\t",eol = "\n", na = "NA", dec = ".", row.names = F, qmethod = c("escape", "double")) + write.table(rld_names, "/home-link/iivow01/git/rnadeseq/error/rld.tsv", append = FALSE, quote = FALSE, sep = "\t",eol = "\n", na = "NA", dec = ".", row.names = F, qmethod = c("escape", "double")) # save table to another variable if pathway analysis if (params$pathway_analysis) { norm_counts <- rld_names rownames(norm_counts) <- norm_counts$Ensembl_ID + write.table(norm_counts, file="/home-link/iivow01/git/rnadeseq/error/norm_countsxy.tsv", sep="\t", quote=F) } } else { # vst transformation + write.table(assays(cds)[["counts"]], file="/home-link/iivow01/git/rnadeseq/error/counts_before_vst.tsv", sep="\t", quote=F) vsd <- vst(cds, blind=FALSE, nsub = params$nsub_genes) vsd_names <- merge(x=gene_names, y=assay(vsd), by.x = "Ensembl_ID", by.y="row.names") write.table(vsd_names, "differential_gene_expression/gene_counts_tables/vst_transformed_gene_counts.tsv", append = FALSE, quote = FALSE, sep = "\t",eol = "\n", na = "NA", dec = ".", row.names = F, qmethod = c("escape", "double")) + write.table(vsd_names, "/home-link/iivow01/git/rnadeseq/error/vsd.tsv", append = FALSE, quote = FALSE, sep = "\t",eol = "\n", na = "NA", dec = ".", row.names = F, qmethod = c("escape", "double")) # save table to another variable if pathway analysis if (params$pathway_analysis) { norm_counts <- vsd_names rownames(norm_counts) <- norm_counts$Ensembl_ID + write.table(norm_counts, file="/home-link/iivow01/git/rnadeseq/error/norm_countsxy.tsv", sep="\t", quote=F) + } } - +write(paste((Sys.time() - now)/60, "min used for transform and bla"), file="/home-link/iivow01/git/rnadeseq/error/times", append=T) +now <- Sys.time() ############################## DIAGNOSTICS AND QUALITY CONTROL PLOTS ############################### # Cooks distances: get important for example when checking knock-out and overexpression studies pdf("differential_gene_expression/plots/further_diagnostics_plots/Cooks-distances.pdf") par(mar=c(10,3,3,3)) -par( mfrow = c(1,2)) +par(mfrow = c(1,2)) boxplot(log10(assays(cds)[["cooks"]]), range=0, las=2,ylim = c(-15, 15),main="log10-Cooks") boxplot(log2(assays(cds)[["cooks"]]), range=0, las=2,ylim = c(-15, 15),main="log2-Cooks") dev.off() @@ -857,6 +873,8 @@ for (i in resultsNames(cds)[-1]) { dev.off() rm(res,qs,bins,ratios,use,h1,h2,colori) } +write(paste((Sys.time() - now)/60, "min used for diag. plots"), file="/home-link/iivow01/git/rnadeseq/error/times", append=T) + ``` @@ -864,6 +882,7 @@ for (i in resultsNames(cds)[-1]) { ```{r summary_stats, echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/")), results = 'asis'} +now <- Sys.time() cat(paste0("*** # Read mapping and assignment @@ -878,27 +897,52 @@ The read mapping and gene assignment statistics, together with other quality con The table below, extracted from the `MultiQC` report, shows a summary of the bioinformatics analysis quality control. Note: Duplicate intercept (%) is the percentage of duplicate reads for the intercept of a linear model of duplicate reads vs read counts. High numbers of duplicates at low read counts can indicate low library complexity with technical duplication." )) -df_DE <- read.table(file = paste0(wd,"/QC/multiqc_data/multiqc_general_stats.txt"), header=TRUE, sep="\t") -is.num <- sapply(df_DE, is.numeric) -df_DE[is.num] <- lapply(df_DE[is.num], round, 2) -columns <- c("Sample", + +if (file.exists(paste0(wd,"/QC/multiqc_data/multiqc_general_stats.txt"))) { + mqc_stats <- read.table(file = paste0(wd,"/QC/multiqc_data/multiqc_general_stats.txt"), header=TRUE, sep="\t") + mqc_version <- 'old_mqc' +} else if (file.exists(paste0(wd,"/QC/multiqc_data/multiqc_samtools_stats.txt"))) { + mqc_stats <- read.table(file = paste0(wd,"/QC/multiqc_data/multiqc_samtools_stats.txt"), header=TRUE, sep="\t") + mqc_version <- 'new_mqc' +} else { + stop("Could not find a suitable multiqc table; please provide a correct multiqc.zip file or omit the parameter altogether") +} + +is.num <- sapply(mqc_stats, is.numeric) +mqc_stats[is.num] <- lapply(mqc_stats[is.num], round, 2) + +if (mqc_version == 'old_mqc') { + columns <- c( + "Sample", "FastQC_mqc.generalstats.fastqc.total_sequences", "FastQC_mqc.generalstats.fastqc.percent_duplicates", "FastQC_mqc.generalstats.fastqc.percent_gc", "Cutadapt_mqc.generalstats.cutadapt.percent_trimmed", "STAR_mqc.generalstats.star.uniquely_mapped_percent", - "featureCounts_mqc.generalstats.featurecounts.percent_assigned") -df_DE_selected <- df_DE[,columns] + "featureCounts_mqc.generalstats.featurecounts.percent_assigned" + ) +} else { + columns <- c( + "Sample", + "raw_total_sequences", + "reads_duplicated_percent", + "reads_mapped_percent" + ) +} +mqc_stats_selected <- mqc_stats[,columns] # The following gets rid of multiQC entries that are spread across two rows, one ending with -fw (see https://github.com/qbic-pipelines/rnadeseq/issues/114) -sum_NA <- function(x) {if (all(is.na(x))) x[NA_integer_] else sum(x, na.rm = TRUE)} -df_DE_selected$Sample <- gsub("-fw", "", df_DE_selected$Sample) -df_DE_selected <- df_DE_selected %>% group_by(Sample) %>% summarise_all(sum_NA) +sum_NA <- function(x) {if (all(is.na(x))) x[NA_integer_] else sum(x, na.rm = TRUE)} # Function to sum across rows if available, else NA +mqc_stats_selected$Sample <- gsub("-fw", "", mqc_stats_selected$Sample) # Remove -fw to produce duplicate rows for summarising +mqc_stats_selected <- mqc_stats_selected %>% group_by(Sample) %>% summarise_all(sum_NA) +# In newer RNASeq versions (after 1.4.2?), duplicate rows might not end with -fw, but with _1 and _2 instead; deal with those in the following lines +avg_NA <- function(x) {if (all(is.na(x))) x[NA_integer_] else mean(x, na.rm = TRUE)} # Function to average across rows if available, else NA +mqc_stats_selected$Sample <- sub("_[0-9]+$", "", mqc_stats_selected$Sample) # Remove _1 etc. to produce duplicate rows for summarising +mqc_stats_selected <- mqc_stats_selected %>% group_by(Sample) %>% summarise_all(avg_NA) -df_DE_selected$Sample <- substr(df_DE_selected$Sample, 1, 10) -colnames(df_DE_selected) <- c("Sample", "C1", "C2", "C3", "C4", "C5", "C6") -n_rows = nrow(df_DE_selected) +mqc_stats_selected$Sample <- substr(mqc_stats_selected$Sample, 1, 10) +n_rows = nrow(mqc_stats_selected) metadata <- read.table((paste0(wd, "/differential_gene_expression/metadata/metadata.tsv")), header=TRUE, sep="\t") metadata = metadata[,c("QBiC.Code")] @@ -906,26 +950,44 @@ metadata = metadata[,c("QBiC.Code")] #the following makes metadata a df again (in R, when extracting only 1 column from a df, it becomes a list) metadata <- data.frame(matrix(unlist(metadata))) colnames(metadata) <- c("Sample") -table_complete = merge(metadata,df_DE_selected,by="Sample") -colnames = c("Sample", "Number of reads (M)", "Duplicates (%)", "GC (%)", "Trimmed reads (%)", "Mapped reads (%)", "Assigned reads (%)") -colnames(table_complete) <- c("Sample", "C1", "C2", "C3", "C4", "C5", "C6") + +table_complete = merge(metadata,mqc_stats_selected,by="Sample") + +if (mqc_version == 'old_mqc') { + colnames = c("Sample", "Number of reads (M)", "Duplicates (%)", "GC (%)", "Trimmed reads (%)", "Mapped reads (%)", "Assigned reads (%)") + colnames(table_complete) <- c("Sample", "ReadNumber", "DuplicateReadsIntercept", "GCcontent", "TrimmedReads", "MappedReads", "AssignedReads") + table_complete <- table_complete %>% + mutate( + Sample = Sample, + ReadNumber = color_bar("lightblue")(round((ReadNumber/1000000),2)), + DuplicateReadsIntercept = ifelse(DuplicateReadsIntercept > 1, + cell_spec(DuplicateReadsIntercept, color="orange", bold=T), + cell_spec(DuplicateReadsIntercept, color="black")), + GCcontent = GCcontent, + TrimmedReads = TrimmedReads, + MappedReads = ifelse(MappedReads > 80, + color_bar("lightblue")(MappedReads), + color_bar("orange")(MappedReads)), + AssignedReads = ifelse(AssignedReads > 60, + color_bar("lightblue")(AssignedReads), + color_bar("orange")(AssignedReads)) + ) +} else { + colnames = c("Sample", "Number of reads (M)", "Duplicates (%)", "Mapped reads (%)") + colnames(table_complete) <- c("Sample", "ReadNumber", "DuplicateReadsIntercept", "MappedReads") + table_complete <- table_complete %>% + mutate( + Sample = Sample, + ReadNumber = color_bar("lightblue")(round((ReadNumber/1000000),2)), + DuplicateReadsIntercept = ifelse(DuplicateReadsIntercept > 1, + cell_spec(DuplicateReadsIntercept, color="orange", bold=T), + cell_spec(DuplicateReadsIntercept, color="black")), + MappedReads = ifelse(MappedReads > 80, color_bar("lightblue")(MappedReads), color_bar("orange")(MappedReads)) + ) +} + +# This does some final formatting of the table in the report table_complete %>% - mutate( - Sample = Sample, - ReadNumber = color_bar("lightblue")(round((C1/1000000),2)), - DuplicateReadsIntercept = ifelse(C2 > 1, - cell_spec(C2, color="orange", bold=T), - cell_spec(C2, color="black")), - GCcontent = C3, - TrimmedReads = C4, - MappedReads = ifelse(C5 > 80, - color_bar("lightblue")(C5), - color_bar("orange")(C5)), - AssignedReads = ifelse(C6 > 60, - color_bar("lightblue")(C6), - color_bar("orange")(C6)) - ) %>% - dplyr::select(1,8:13) %>% kable(escape=F,row.names=F, col.names=colnames) %>% kable_styling("hover") %>% scroll_box(width = "100%", height = "400px") @@ -948,7 +1010,7 @@ The read mapping statistics to the reference genome for each sample are shown be ```{r STAR_percentages_plot, echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/")), out.width="160%", dpi=1800, fig.cap="STAR: Mapping Statistics", fig.align='center'} # Disable auto-conversion of paths to relative paths as we provide include_graphics() with absolute paths. Also, error=F is necessary in every include_graphic() call to not fail with an error knitr.graphics.rel_path = F -knitr::include_graphics(paste0(wd, "/QC/multiqc_plots/svg/mqc_star_alignment_plot_1_pc.svg"), error=F) +knitr::include_graphics(paste0(wd, "/QC/multiqc_plots/svg/", ifelse(mqc_version == 'old_mqc',"mqc_star_alignment_plot_1_pc.svg", "mqc_samtools_alignment_plot_1_pc.svg")), error=F) ``` ```{r STAR_readnums,echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/"))} @@ -960,14 +1022,15 @@ cat(paste0("*** ``` ```{r STAR_readnums_plot, echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/")), out.width="160%", dpi=1800, fig.cap="STAR: Mapping Statistics", fig.align='center'} -knitr::include_graphics(paste0(wd, "/QC/multiqc_plots/svg/mqc_star_alignment_plot_1.svg"), error=F) +knitr::include_graphics(paste0(wd, "/QC/multiqc_plots/svg/", ifelse(mqc_version == 'old_mqc',"mqc_star_alignment_plot_1.svg", "mqc_samtools_alignment_plot_1.svg")), error=F) + ``` ```{r FC_read_assignment, echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/"))} cat(paste0("*** \ - +", ifelse(mqc_version == 'old_mqc', " ## Read assignment statistics {.tabset} The statistics of read assignment to genes are shown below. Most reads should be assigned. Interpretation of the different assignment types: @@ -982,14 +1045,18 @@ The statistics of read assignment to genes are shown below. Most reads should be ### Percentages {-}
-")) +", " +## Read alignment statistics {.tabset} + +The statistics of read alignment to genes are shown below for pipeline runs on data produced by `rnaseq <= v1.4.2`. If a later version was used, these plots are not generated by rnaseq anymore. However, you can find `star/featurecounts/.featureCounts.txt.summary` or `star_salmon/featurecounts/.featureCounts.txt.summary` files in the rnaseq output folder which contain the necessary information for creating such plots. +"))) ``` -```{r FC_percentages_plot, echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/")), out.width="160%", dpi=1200, fig.cap="featureCounts: Assignments", fig.align='center'} +```{r FC_percentages_plot, echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/")) && mqc_version == 'old_mqc', out.width="160%", dpi=1200, fig.cap=ifelse(mqc_version == 'old_mqc', "featureCounts: Assignments", "STAR: Alignments"), fig.align='center'} knitr::include_graphics(paste0(wd, "/QC/multiqc_plots/svg/mqc_featureCounts_assignment_plot_1_pc.svg"), error=F) ``` -```{r FC_readnums, echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/"))} +```{r FC_readnums, echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/")) && mqc_version == 'old_mqc'} cat(paste0("***
@@ -999,7 +1066,7 @@ cat(paste0("*** ")) ``` -```{r FC_readnums_plot, echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/")), out.width="160%", dpi=1200, fig.cap="featureCounts: Assignments", fig.align='center'} +```{r FC_readnums_plot, echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/")) && mqc_version == 'old_mqc', out.width="160%", dpi=1200, fig.cap=ifelse(mqc_version == 'old_mqc', "featureCounts: Assignments", "STAR: Alignments"), fig.align='center'} knitr::include_graphics(paste0(wd, "/QC/multiqc_plots/svg/mqc_featureCounts_assignment_plot_1.svg"), error=F) ``` @@ -1037,11 +1104,30 @@ cat(paste0("*** knitr::include_graphics(paste0(wd, "/QC/multiqc_plots/svg/mqc_rseqc_read_distribution_plot_1.svg"), error=F) ``` +```{r Read_strand, echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/"))} +cat(paste0("*** + +\ + +## Read strandedness {.tabset} + +The following plots show the number/percentage of reads that match the strandedness of overlapping transcripts. It can be used to infer whether RNA-seq library preps are stranded (sense or antisense). + +
+")) +``` + +```{r Read_strandpercentages_plot, echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/")), out.width="160%", dpi=1200, fig.cap="RSeQC: Read Distribution", fig.align='center'} +knitr::include_graphics(paste0(wd, "/QC/multiqc_plots/svg/mqc_rseqc_infer_experiment_plot_1.svg"), error=F) +``` + ```{r MQC_end, echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/"))} cat(paste0("***
\ ")) +write(paste((Sys.time() - now)/60, "min used for mqc"), file="/home-link/iivow01/git/rnadeseq/error/times", append=T) + ``` @@ -1052,7 +1138,7 @@ cat(paste0("*** The raw count table and normalized count tables are available [here](./differential_gene_expression/gene_counts_tables). The differential expression analysis is performed using the raw gene count table. -For PCA analysis and heatmap plotting, the `r norm_method_long_text` normalized gene counts were used. +For PCA analysis and heatmap plotting, the `r norm_method_long_text` normalized gene counts were used. `r norm_method_override_text` `r nsub_genes_text` @@ -1065,6 +1151,7 @@ The original plot can be downloaded [here](./differential_gene_expression/plots/ \ ```{r PCA_ifNoBatch_plot, echo=F, message=F, warning=F, out.width="80%", dpi=1200, fig.align='center'} +now <- Sys.time() ############################ PCA PLOTS ######################## pcaData <- plotPCA(if (run_rlog) rld else vsd,intgroup=c("combfactor"), ntop=dim(if (run_rlog) rld else vsd)[1], returnData=TRUE) percentVar <- round(100*attr(pcaData, "percentVar")) @@ -1156,6 +1243,8 @@ Two samples have more similar gene expression if their distance is closer to 0 ( The original plot can be downloaded [here](./differential_gene_expression/plots/Heatmaps_of_distances.pdf). ```{r heatmap_calculate, include=FALSE} +write(paste((Sys.time() - now)/60, "min used for PCA"), file="/home-link/iivow01/git/rnadeseq/error/times", append=T) + ################## SAMPLE DISTANCES HEATMAP ################## # Sample distances, this chunk is hidden as otherwise, the heatmap is shown multiple times sampleDists <- dist(t(assay(if (run_rlog) rld else vsd))) @@ -1225,7 +1314,7 @@ These comparisons are called _contrasts_. For this analysis, the following contr ```{r contrast_calc, echo=FALSE, message=FALSE, warning=FALSE, results = 'hide'} - +now <- Sys.time() # Contrasts coefficient table write in metadata coefficients <- resultsNames(cds) coef_tab <- data.frame(coef=coefficients) @@ -1251,7 +1340,7 @@ if (isProvided(params$path_contrast_matrix)){ # Check that contrast matrix is valid if (length(coefficients) != nrow(contrasts)){ - stop("Error: Your contrast table has a different number of rows than the number of coefficients in the DESeq2 model.") + stop("Your contrast table has a different number of rows than the number of coefficients in the DESeq2 model.") } ## Contrast calculation for contrast matrix @@ -1348,6 +1437,7 @@ if (isProvided(params$path_contrast_list)) { contrast_names <- append(contrast_names, contname) } } + if (isProvided(params$path_contrast_pairs)) { contrasts <- read.table(path_contrast_pairs, sep="\t", header = T, colClasses = "character") write.table(contrasts, file="differential_gene_expression/metadata/contrast_pairs.tsv", sep="\t", quote=F, col.names = T, row.names = F) @@ -1488,6 +1578,8 @@ if (isProvided(params$round_DE) && params$round_DE>-1) { #write to file write.table(DE_genes_final_table, "differential_gene_expression/final_gene_table/final_DE_gene_list.tsv", append = FALSE, quote = FALSE, sep = "\t",eol = "\n", na = "NA", dec = ".", row.names = F, col.names = T, qmethod = c("escape", "double")) +write(paste((Sys.time() - now)/60, "min used for contrast calc"), file="/home-link/iivow01/git/rnadeseq/error/times", append=T) + ``` @@ -1583,6 +1675,7 @@ config(pg, modeBarButtonsToAdd = list(svg_exp)) %>% layout(margin = list(l=80, b ``` ```{r box_plots_noBatch, echo=F, message=F} +now <- Sys.time() # Decide which object to use for boxplots without batch correction if (run_rlog) { box_object <- rld_noncorrected @@ -1650,7 +1743,7 @@ if (isProvided(params$path_genelist)) { ggsave(filename=paste("differential_gene_expression/plots/boxplots_requested_genes/",requested_genes_plot_gene_name[i],"_",requested_genes_plot_Ensembl[i],".svg",sep=""), width=10, height=5, plot=plot) ggsave(filename=paste("differential_gene_expression/plots/boxplots_requested_genes/",requested_genes_plot_gene_name[i],"_",requested_genes_plot_Ensembl[i],".png",sep=""), width=10, height=5, plot=plot) ggsave(filename=paste("differential_gene_expression/plots/boxplots_requested_genes/",requested_genes_plot_gene_name[i],"_",requested_genes_plot_Ensembl[i],".pdf",sep=""), width=10, height=5, plot=plot) - } + } } ``` @@ -1717,7 +1810,7 @@ if (isProvided(params$path_genelist)) { ggsave(filename=paste0("differential_gene_expression/plots/boxplots_requested_genes/",requested_genes_plot_gene_name[i],"_",requested_genes_plot_Ensembl[i],"_after_batchcorrect.svg"), width=10, height=5, plot=plot) ggsave(filename=paste0("differential_gene_expression/plots/boxplots_requested_genes/",requested_genes_plot_gene_name[i],"_",requested_genes_plot_Ensembl[i],"_after_batchcorrect.png"), width=10, height=5, plot=plot) ggsave(filename=paste0("differential_gene_expression/plots/boxplots_requested_genes/",requested_genes_plot_gene_name[i],"_",requested_genes_plot_Ensembl[i],"_after_batchcorrect.pdf"), width=10, height=5, plot=plot) - } + } } ``` @@ -1734,6 +1827,8 @@ if (!inherits(df_gene, 'try-error')){ names = rownames(df_gene) gene = TRUE } +write(paste((Sys.time() - now)/60, "min used for boxplot stuff"), file="/home-link/iivow01/git/rnadeseq/error/times", append=T) +nownow <- Sys.time() ``` ```{r genes_block, echo=FALSE, results='asis', eval=gene} @@ -1746,7 +1841,7 @@ cat("Plots for the [genes of interest](./differential_gene_expression/metadata/g ```{r load_PA_libs, eval=params$pathway_analysis, echo=FALSE, message=FALSE, warning=FALSE, results = 'hide'} - +now <- Sys.time() # Load libs invisibly invisible(lapply(c( "gprofiler2", @@ -1771,8 +1866,12 @@ BiocManager::install(params$species_library, lib=species_dir, version="3.17", fo library(params$species_library, lib.loc=species_dir, character.only=T) species_library_installed <- get(params$species_library) -# gprofiler pathway / term sources parameters -datasources <- c("KEGG", "REAC") +# If provided, prepare datasources from comma-separated string +if (isProvided(params$datasources)) { + datasources <- unlist(strsplit(params$datasources, ',')) +} else { + datasources <- c("KEGG", "REAC") +} # Default to 1 for the nb of genes DE in a pathway min_DEG_pathway <- as.integer(params$min_DEG_pathway) @@ -1802,7 +1901,18 @@ mv_status <- F if (isProvided(params$custom_gmt)){ # If custom GMT file was provided, filter it before uploading to use in gost queries (gprofiler will NOT filter itself, so otherwise the output will contain all kinds of datasources, not just the selected ones) - out_gmt <- (Filter(function(line) any(startsWith(line, datasources)), readLines(params$custom_gmt))) + out_gmt <- readLines(params$custom_gmt) + + # If datasources are also provided, filter the GMT + if (isProvided(params$datasources)) { + go_positions <- grep("^GO", datasources) + if (length(go_positions)) { + datasources <- my_vector[-go_positions] # If there are any datasources starting with GO, remove them (gets rid of GO:BP etc.) + datasources <- append(datasources, "GO") # Then add the single entry GO, as otherwise, the filtering might not recognize these datasources + } + out_gmt <- (Filter(function(line) any(startsWith(line, datasources)), out_gmt)) # Remove GMT lines that don't start with a datasource + } + out_path <- paste0(tools::file_path_sans_ext(basename(params$custom_gmt)), "_filtered.gmt") writeLines(out_gmt, out_path) gost_id <- upload_GMT_file(out_path) @@ -1844,6 +1954,11 @@ if (mv_status) { } } +# Set datasources to NULL instead of c() to ensure that the gost queries work +if (!isProvided(params$datasources)) { + datasources <- NULL +} + # ------------------ # Set default params # ------------------ @@ -1859,11 +1974,12 @@ theme_set(theme_classic()) for (file in contrast_files){ - #Reading DE genes list + # Reading DE genes list fname <- tools::file_path_sans_ext(basename(file)) dir.create(paste("pathway_analysis", fname, sep="/")) dir.create(paste("pathway_analysis", fname, "KEGG_pathways", sep="/")) + dir.create(paste("pathway_analysis", fname, "enrichment_plots", sep="/")) DE_genes <- read.csv(file = paste0(path_contrasts, file), sep="\t", header = T) DE_genes <- as.data.frame(DE_genes) @@ -1875,6 +1991,7 @@ for (file in contrast_files){ # Define list of Ensemble IDs (q) to run the pathway analysis q = as.character(DE_genes$Ensembl_ID) + write(paste((Sys.time() - nownow)/60, "min used for weird slow stuff"), file="/home-link/iivow01/git/rnadeseq/error/times", append=T) # gost query if (isProvided(params$set_background)) { @@ -1888,11 +2005,11 @@ for (file in contrast_files){ custom_bg=custom_background, domain_scope="custom_annotated" ) - + pathway_gostres <- gostres$result pathway_gostres <- as.data.frame(pathway_gostres[which(pathway_gostres$significant==TRUE),]) pathway_gostres <- pathway_gostres[which(pathway_gostres$intersection_size>=min_DEG_pathway),] - + # Also run without bg for Venn diagrams gostres_nobg <- gost(query=q, organism=gost_id, @@ -1919,7 +2036,6 @@ for (file in contrast_files){ ) } - # Make data frame of gost result pathway_gostres <- gostres$result @@ -1938,13 +2054,13 @@ for (file in contrast_files){ # Generate non-interactive pathway dotplots in the folder pg <- gostplot(gostres, capped=T, interactive=F) - ggsave(pg, filename = paste0("pathway_analysis", "/", fname, "_gost_pathway_enrichment_plot.pdf"), + ggsave(pg, filename = paste0("pathway_analysis/", "/", fname, "/gost_pathway_gostplot.pdf"), device="pdf", height=10, width=15, units="cm", limitsize=F) - ggsave(pg, filename = paste0("pathway_analysis", "/", fname, "_gost_pathway_enrichment_plot.png"), + ggsave(pg, filename = paste0("pathway_analysis/", "/", fname, "/gost_pathway_gostplot.png"), device="png", height=10, width=15, units="cm", dpi=300, limitsize=F) - ggsave(pg, filename = paste0("pathway_analysis", "/", fname, "_gost_pathway_enrichment_plot.svg"), + ggsave(pg, filename = paste0("pathway_analysis/", "/", fname, "/gost_pathway_gostplot.svg"), device="svg", height=10, width=15, units="cm", dpi=300, limitsize=F) @@ -1958,9 +2074,9 @@ for (file in contrast_files){ no_background=pathway_gostres_nobg$term_name ) pv <- ggvenn::ggvenn(VennInput) - ggsave(paste0("pathway_analysis", "/", fname, "_gost_pathway_venn_diagram.pdf"),pv, device="pdf", width=10, height=15) - ggsave(paste0("pathway_analysis", "/", fname, "_gost_pathway_venn_diagram.png"),pv, device="png", width=10, height=15) - ggsave(paste0("pathway_analysis", "/", fname, "_gost_pathway_venn_diagram.svg"),pv, device="svg", width=10, height=15) + ggsave(paste0("pathway_analysis", "/", fname, "/gost_pathway_venn_diagram.pdf"),pv, device="pdf", width=10, height=15) + ggsave(paste0("pathway_analysis", "/", fname, "/gost_pathway_venn_diagram.png"),pv, device="png", width=10, height=15) + ggsave(paste0("pathway_analysis", "/", fname, "/gost_pathway_venn_diagram.svg"),pv, device="svg", width=10, height=15) } } @@ -1970,7 +2086,7 @@ for (file in contrast_files){ # Save pathway enrichment table in tsv format write.table(pathway_gostres_table, - file = paste0("pathway_analysis", "/", fname, "/", fname, "_pathway_enrichment_results.tsv"), + file = paste0("pathway_analysis", "/", fname, "/pathway_enrichment_results.tsv"), sep="\t", quote = F, col.names = T, row.names = F) print("------------------------------------") @@ -1992,7 +2108,7 @@ for (file in contrast_files){ # Plotting results for df df_subset <- data.frame(Pathway_name = df$short_name, Pathway_code = df$term_id, DE_genes = df$intersection_size, Pathway_size = df$term_size, Fraction_DE = (df$intersection_size / df$term_size), Padj = df$p_value, DE_genes_names = df$intersection) write.table(df_subset, - file = paste0("pathway_analysis", "/", fname, "/", fname, "_", db_source, "_pathway_enrichment_results.tsv"), + file = paste0("pathway_analysis", "/", fname, "/", make.names(db_source), "_pathway_enrichment_results.tsv"), sep="\t", quote = F, col.names = T, row.names = F) # Enriched pathways horizontal barplots of padj values @@ -2004,13 +2120,14 @@ for (file in contrast_files){ scale_fill_continuous(high = "#132B43", low = "#56B1F7") + ggtitle("Enriched pathways") + xlab("") + ylab("Gene fraction (DE genes / Pathway size)") - ggsave(p, filename = paste0("pathway_analysis", "/", fname, "/", fname, "_", db_source, "_pathway_enrichment_plot.pdf"), device = "pdf", height = 5+0.5*nrow(df_subset), units = "cm", limitsize=F) - ggsave(p, filename = paste0("pathway_analysis", "/", fname, "/", fname,"_", db_source, "_pathway_enrichment_plot.png"), device = "png", height = 5+0.5*nrow(df_subset), units = "cm", dpi = 300, limitsize=F) - ggsave(p, filename = paste0("pathway_analysis", "/", fname, "/", fname,"_", db_source, "_pathway_enrichment_plot.svg"), device = "svg", height = 5+0.5*nrow(df_subset), units = "cm", dpi = 300, limitsize=F) + ggsave(p, filename = paste0("pathway_analysis", "/", fname, "/enrichment_plots/", make.names(db_source), "_pathway_enrichment_plot.pdf"), device = "pdf", height = 5+0.5*nrow(df_subset), units = "cm", limitsize=F) + ggsave(p, filename = paste0("pathway_analysis", "/", fname, "/enrichment_plots/", make.names(db_source), "_pathway_enrichment_plot.png"), device = "png", height = 5+0.5*nrow(df_subset), units = "cm", limitsize=F) + ggsave(p, filename = paste0("pathway_analysis", "/", fname, "/enrichment_plots/", make.names(db_source), "_pathway_enrichment_plot.svg"), device = "svg", height = 5+0.5*nrow(df_subset), units = "cm", limitsize=F) # Plotting heatmaps and KEGG pathways for all pathways print("Plotting heatmaps...") if (nrow(df) <= 100 & nrow(df) > 0) { + capture.output(df, file="/home-link/iivow01/git/rnadeseq/error/df.tsv", sep="\t", quote=F) conditions <- grepl("Condition", colnames(metadata)) metadata_save <- as.data.frame(metadata[,conditions]) metadata_name <- metadata[,c("QBiC.Code", "Secondary.Name")] @@ -2018,8 +2135,15 @@ for (file in contrast_files){ for (i in c(1:nrow(df))){ pathway <- df[i,] + capture.output(pathway, file="/home-link/iivow01/git/rnadeseq/error/pathway.tsv", sep="\t", quote=F) + gene_list <- unlist(strsplit(pathway$intersection, ",")) + write(paste((Sys.time() - now)/60, "min used for PA before mat stuff"), file="/home-link/iivow01/git/rnadeseq/error/times", append=T) + + capture.output(gene_list, file="/home-link/iivow01/git/rnadeseq/error/gene_list") mat <- norm_counts[gene_list, ] + write.table(mat, file="/home-link/iivow01/git/rnadeseq/error/mat.tsv", sep="\t", quote=F) + write.table(norm_counts, file="/home-link/iivow01/git/rnadeseq/error/norm_countsss.tsv", sep="\t", quote=F) rownames(mat) <- mat$gene_name mat$gene_name <- NULL mat <- data.matrix(mat) @@ -2027,38 +2151,39 @@ for (file in contrast_files){ # Skip the heatmaps for CI tests because especially png() works only very unreliably for the heatmaps for some reason if (!isProvided(params$citest) & nrow(mat)>1){ dir.create(paste("pathway_analysis", fname, "pathway_heatmaps", sep="/")) - - # Because the image devices sometimes don't work here, surround them with a tryCatch so that the code continues even if a certain heatmap is not saved + + # Because the image devices sometimes don't work here, surround them with a tryCatch so that the code continues even if a certain heatmap is not saved tryCatch( { - png(filename = paste0("pathway_analysis", "/",fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_",fname, ".png"), width = ifelse(nrow(mat)<=20, 200, 125)+ncol(mat)*40, height = 50+nrow(mat)*50) + png(filename = paste0("pathway_analysis", "/",fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", make.names(pathway$term_id), ".png"), width = ifelse(nrow(mat)<=20, 200, 125)+ncol(mat)*40, height = 50+nrow(mat)*50) pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T) dev.off() }, error=function(e) { - print(paste0("Could not save Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_",fname, ".png because of the following error:\n", e)) + print(paste0("Could not save pathway_analysis", "/",fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", make.names(pathway$term_id), ".png because of the following error:\n", e)) } ) tryCatch( { - pdf(paste0("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".pdf"), width = ifelse(nrow(mat)<=20, 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005) + pdf(paste0("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", make.names(pathway$term_id), ".pdf"), width = ifelse(nrow(mat)<=20, 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005) pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T) dev.off() }, error=function(e) { - print(paste0("Could not save Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".pdf because of the following error:\n", e)) + print(paste0("Could not save pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", make.names(pathway$term_id), ".pdf because of the following error:\n", e)) } ) tryCatch( { - svg(paste0("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".svg"), width = ifelse(nrow(mat)<=20, 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005) + svg(paste0("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", make.names(pathway$term_id), ".svg"), width = ifelse(nrow(mat)<=20, 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005) pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T) dev.off() }, error=function(e) { - print(paste0("Could not save Heatmap_normalized_counts_", pathway$source, "_", pathway$term_id, "_", fname, ".svg because of the following error:\n", e)) + print(paste0("Could not save pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", make.names(pathway$term_id), ".svg because of the following error:\n", e)) } ) + graphics.off() # Close all still open devices from errors. Tried doing this in finally, but that doesn't work } # Plotting pathway view only for kegg pathways @@ -2070,9 +2195,7 @@ for (file in contrast_files){ print_string <- paste0("Plotting pathway: ", pathway_kegg) gene.data = DE_genes gene.data.subset = gene.data[gene.data$Ensembl_ID %in% gene_list, c("Ensembl_ID","log2FoldChange")] - entrez_ids = AnnotationDbi::mapIds(species_library_installed, keys=as.character(gene.data.subset$Ensembl_ID), column = "ENTREZID", keytype=params$keytype, multiVals="first") - gene.data.subset <- gene.data.subset[!(is.na(entrez_ids)),] if (length(entrez_ids)!=length(unique(entrez_ids))) { @@ -2128,6 +2251,7 @@ if (isProvided(params$path_genelist)){ rownames(norm_counts) <- norm_counts$Ensembl_ID mat <- norm_counts[gene_list, ] + rownames(mat) <- mat$gene_name mat$gene_name <- NULL mat$Ensembl_ID <- NULL @@ -2170,9 +2294,11 @@ Inside the pathway analysis results folder, a subfolder for each contrast used f

Expand/collapse - `*_gost_pathway_venn_diagram.pdf/png` - - Venn diagrams showing the numbers of enriched pathways when using a background gene list vs when not using a bg list. -- `*_KEGG_pathway_enrichment_plot.pdf/png` - - Barplots showing the proportion of differentially expressed genes in the pathway. + - Venn diagrams showing the numbers of enriched pathways when using a background gene list vs when not using a bg list. +- `enrichment_plots/*_pathway_enrichment_plot.{pdf/png/svg}` + - Barplots showing the proportion of differentially expressed genes in the pathway for a certain pathway database. +- `gost_pathway_gostplot.{pdf/png/svg}` + - Manhattan plots displaying all enriched pathways. - `KEGG_pathways/` - Contains the KEGG pathways graphs with the log fold change of the differentially expressed genes. - `pathway_heatmaps` @@ -2211,7 +2337,7 @@ names(q_list) <- q_names if (length(q_list) > 0) { #gost query - + if (isProvided(params$set_background)) { gostres <- gost(query=q_list, organism=gost_id, @@ -2288,6 +2414,8 @@ You can find these heatmaps inside the [pathway_analysis](./pathway_analysis) fo and the subfolder: `pathway_heatmaps`. \ ")) +write(paste((Sys.time() - now)/60, "min used for all PA stuff"), file="/home-link/iivow01/git/rnadeseq/error/times", append=T) + ``` @@ -2422,7 +2550,7 @@ as.character(params$revision), "` pipeline [^5], which was written using the nf-core template [@ewels2020nf]. For differential expression analysis, the read quantification data resulting from `", quant_tool, "` were processed with the R package `DESeq2 v", packageVersion("DESeq2"), -"` [@love2014differential]. The thresholds for differentially expressed genes were set to ", +"` [@love2014differential]. The thresholds for differentially expressed genes were set to ", pval_text, " for the p-value and ", logFC_text, " for the log2 Fold Change." )) @@ -2434,16 +2562,25 @@ name_species = params$species_library version_species = packageVersion(name_species) if (isProvided(params$custom_gmt)){ - database_string <- paste0("Also, a gprofiler query with this custom GMT file was done: `", basename(params$custom_gmt), "`. Entries of the following datasources were considered: ") - dbs <- paste(datasources, collapse="`, `") - database_string <- paste0(database_string, "`", dbs, "`.") -} else { + database_string <- paste0("Also, a gprofiler query with this custom GMT file was done: `", basename(params$custom_gmt), "`.") + + if (isProvided(params$datasources)) { + database_string <- paste(database_string, "Entries of the following datasources were considered: ") + dbs <- paste(datasources, collapse="`, `") + database_string <- paste0(database_string, "`", dbs, "`.") + } else { + database_string <- "" + } + +} else if (isProvided(params$datasources)) { database_string = "Also, the following databases were queried: " database_version_info <- get_version_info(params$organism) database_string <- paste0(database_string, "`gprofiler: ", database_version_info$gprofiler_version, "`") for (db in datasources) { database_string <- paste0(database_string, ", `", db, ": ", database_version_info$sources[[db]]$version, "`") } +} else { + database_string <- paste("All datasources available in g:Profiler were considered (check https://biit.cs.ut.ee/gprofiler/gost and https://biit.cs.ut.ee/gprofiler/page/docs for more info).") } cat(paste0( @@ -2482,4 +2619,4 @@ sessioninfo::session_info()%>% [^6]: https://github.com/nf-core/smrnaseq [^7]: https://github.com/OpenGene/fastp -[^8]: https://github.com/friedlanderlab/mirtrace +[^8]: https://github.com/friedlanderlab/mirtrace \ No newline at end of file diff --git a/environment.yml b/environment.yml index cf3dc64d..cede896c 100644 --- a/environment.yml +++ b/environment.yml @@ -1,7 +1,7 @@ # You can use this file to create a conda environment for this pipeline: # conda env create -f environment.yml # use this to find packages: https://anaconda.org/ -name: qbic-pipelines-rnadeseq-2.2 +name: qbic-pipelines-rnadeseq-dev channels: - bioconda - conda-forge diff --git a/modules/local/report.nf b/modules/local/report.nf index eac509d0..167fceb5 100644 --- a/modules/local/report.nf +++ b/modules/local/report.nf @@ -1,6 +1,6 @@ process REPORT { - container 'ghcr.io/qbic-pipelines/rnadeseq:2.2' + container 'ghcr.io/qbic-pipelines/rnadeseq:dev' input: path gene_counts From 00bdd7522ed0c56e27b6cd2225757f450ee1b538 Mon Sep 17 00:00:00 2001 From: WackerO Date: Fri, 12 Apr 2024 15:24:36 +0200 Subject: [PATCH 28/73] Reintroduced changes after finding previous error --- assets/RNAseq_report.Rmd | 53 ++++-------------------- bin/Execute_report.R | 2 + conf/test.config | 1 + conf/test_custom_gmt.config | 1 + conf/test_smrnaseq.config | 1 + conf/test_star_rsem.config | 3 +- conf/test_star_salmon.config | 1 + docs/output.md | 6 ++- docs/usage.md | 4 +- modules/local/report.nf | 5 ++- nextflow.config | 3 +- nextflow_schema.json | 6 ++- tests/test.yml | 49 +++++++++++----------- tests/test_batcheffect.yml | 0 tests/test_contrast_list.yml | 0 tests/test_contrast_matrix.yml | 0 tests/test_custom_gmt.yml | 12 ++++-- tests/test_full.yml | 36 ++++++++-------- tests/test_no_multiqc.yml | 0 tests/test_relevel.yml | 0 tests/test_smrnaseq.yml | 28 ++++++------- tests/test_star_rsem.yml | 75 ++++++++++++++++++---------------- tests/test_star_salmon.yml | 8 ++-- tests/test_use_vst.yml | 0 24 files changed, 141 insertions(+), 153 deletions(-) mode change 100644 => 100755 tests/test.yml mode change 100644 => 100755 tests/test_batcheffect.yml mode change 100644 => 100755 tests/test_contrast_list.yml mode change 100644 => 100755 tests/test_contrast_matrix.yml mode change 100644 => 100755 tests/test_custom_gmt.yml mode change 100644 => 100755 tests/test_full.yml mode change 100644 => 100755 tests/test_no_multiqc.yml mode change 100644 => 100755 tests/test_relevel.yml mode change 100644 => 100755 tests/test_smrnaseq.yml mode change 100644 => 100755 tests/test_star_rsem.yml mode change 100644 => 100755 tests/test_star_salmon.yml mode change 100644 => 100755 tests/test_use_vst.yml diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index 3cd6ad23..921dbf1b 100644 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -349,7 +349,6 @@ Auf der Morgenstelle 10, 72076 Tuebingen ```{r read_metadata, echo=FALSE, message=FALSE, warning=FALSE, results='asis'} -now <- Sys.time() # Load metadata: sample preparations tsv file from qPortal metadata <- read.table(params$path_metadata, sep="\t", header=TRUE,na.strings =c("","NaN"), quote=NULL, stringsAsFactors=F, dec=".", fill=TRUE, row.names=1) if (params$pathway_analysis) { @@ -496,7 +495,6 @@ if(file.exists(QAfile)){ } else { no_RIN <- TRUE } -write(paste((Sys.time() - now)/60, "min used for read_metadata"), file="/home-link/iivow01/git/rnadeseq/error/times", append=F) ``` ```{r RIN_block, echo=FALSE, results='asis', eval=RIN} @@ -525,7 +523,7 @@ DT::datatable(df_QA_neat, options = list(scrollX = "100%", scrollY = "400px", p ```{r DESeq, echo=FALSE, message=FALSE, warning=FALSE, results = 'hide'} -now <- Sys.time() + ################## RUN DESEQ2 ###################################### # Run DESeq function if (params$input_type %in% c("featurecounts", "smrnaseq")) { @@ -639,8 +637,6 @@ if (isProvided(params$path_relevel)) { # Run actual DESeq analysis cds <- DESeq(cds) -write(paste((Sys.time() - now)/60, "min used for run DESeq2"), file="/home-link/iivow01/git/rnadeseq/error/times", append=T) -now <- Sys.time() # sizeFactors(cds) as indicator of library sequencing depth if (params$input_type %in% c("featurecounts", "smrnaseq")) { sf <- sizeFactors(cds) @@ -699,38 +695,30 @@ norm_method_text <- ifelse(run_rlog, "rlog", "vst") if (run_rlog){ # rlog transformation - write.table(assays(cds)[["counts"]], file="/home-link/iivow01/git/rnadeseq/error/counts_before_rlog.tsv", sep="\t", quote=F) rld <- rlog(cds, blind=FALSE) rld_names <- merge(x=gene_names, y=assay(rld), by.x = "Ensembl_ID", by.y="row.names") write.table(rld_names, "differential_gene_expression/gene_counts_tables/rlog_transformed_gene_counts.tsv", append = FALSE, quote = FALSE, sep = "\t",eol = "\n", na = "NA", dec = ".", row.names = F, qmethod = c("escape", "double")) - write.table(rld_names, "/home-link/iivow01/git/rnadeseq/error/rld.tsv", append = FALSE, quote = FALSE, sep = "\t",eol = "\n", na = "NA", dec = ".", row.names = F, qmethod = c("escape", "double")) # save table to another variable if pathway analysis if (params$pathway_analysis) { norm_counts <- rld_names rownames(norm_counts) <- norm_counts$Ensembl_ID - write.table(norm_counts, file="/home-link/iivow01/git/rnadeseq/error/norm_countsxy.tsv", sep="\t", quote=F) } } else { # vst transformation - write.table(assays(cds)[["counts"]], file="/home-link/iivow01/git/rnadeseq/error/counts_before_vst.tsv", sep="\t", quote=F) vsd <- vst(cds, blind=FALSE, nsub = params$nsub_genes) vsd_names <- merge(x=gene_names, y=assay(vsd), by.x = "Ensembl_ID", by.y="row.names") write.table(vsd_names, "differential_gene_expression/gene_counts_tables/vst_transformed_gene_counts.tsv", append = FALSE, quote = FALSE, sep = "\t",eol = "\n", na = "NA", dec = ".", row.names = F, qmethod = c("escape", "double")) - write.table(vsd_names, "/home-link/iivow01/git/rnadeseq/error/vsd.tsv", append = FALSE, quote = FALSE, sep = "\t",eol = "\n", na = "NA", dec = ".", row.names = F, qmethod = c("escape", "double")) # save table to another variable if pathway analysis if (params$pathway_analysis) { norm_counts <- vsd_names rownames(norm_counts) <- norm_counts$Ensembl_ID - write.table(norm_counts, file="/home-link/iivow01/git/rnadeseq/error/norm_countsxy.tsv", sep="\t", quote=F) - } } -write(paste((Sys.time() - now)/60, "min used for transform and bla"), file="/home-link/iivow01/git/rnadeseq/error/times", append=T) -now <- Sys.time() + ############################## DIAGNOSTICS AND QUALITY CONTROL PLOTS ############################### # Cooks distances: get important for example when checking knock-out and overexpression studies pdf("differential_gene_expression/plots/further_diagnostics_plots/Cooks-distances.pdf") @@ -873,8 +861,6 @@ for (i in resultsNames(cds)[-1]) { dev.off() rm(res,qs,bins,ratios,use,h1,h2,colori) } -write(paste((Sys.time() - now)/60, "min used for diag. plots"), file="/home-link/iivow01/git/rnadeseq/error/times", append=T) - ``` @@ -882,7 +868,6 @@ write(paste((Sys.time() - now)/60, "min used for diag. plots"), file="/home-link ```{r summary_stats, echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/")), results = 'asis'} -now <- Sys.time() cat(paste0("*** # Read mapping and assignment @@ -1126,8 +1111,6 @@ cat(paste0("*** \ ")) -write(paste((Sys.time() - now)/60, "min used for mqc"), file="/home-link/iivow01/git/rnadeseq/error/times", append=T) - ``` @@ -1151,7 +1134,6 @@ The original plot can be downloaded [here](./differential_gene_expression/plots/ \ ```{r PCA_ifNoBatch_plot, echo=F, message=F, warning=F, out.width="80%", dpi=1200, fig.align='center'} -now <- Sys.time() ############################ PCA PLOTS ######################## pcaData <- plotPCA(if (run_rlog) rld else vsd,intgroup=c("combfactor"), ntop=dim(if (run_rlog) rld else vsd)[1], returnData=TRUE) percentVar <- round(100*attr(pcaData, "percentVar")) @@ -1243,8 +1225,6 @@ Two samples have more similar gene expression if their distance is closer to 0 ( The original plot can be downloaded [here](./differential_gene_expression/plots/Heatmaps_of_distances.pdf). ```{r heatmap_calculate, include=FALSE} -write(paste((Sys.time() - now)/60, "min used for PCA"), file="/home-link/iivow01/git/rnadeseq/error/times", append=T) - ################## SAMPLE DISTANCES HEATMAP ################## # Sample distances, this chunk is hidden as otherwise, the heatmap is shown multiple times sampleDists <- dist(t(assay(if (run_rlog) rld else vsd))) @@ -1314,7 +1294,7 @@ These comparisons are called _contrasts_. For this analysis, the following contr ```{r contrast_calc, echo=FALSE, message=FALSE, warning=FALSE, results = 'hide'} -now <- Sys.time() + # Contrasts coefficient table write in metadata coefficients <- resultsNames(cds) coef_tab <- data.frame(coef=coefficients) @@ -1578,8 +1558,6 @@ if (isProvided(params$round_DE) && params$round_DE>-1) { #write to file write.table(DE_genes_final_table, "differential_gene_expression/final_gene_table/final_DE_gene_list.tsv", append = FALSE, quote = FALSE, sep = "\t",eol = "\n", na = "NA", dec = ".", row.names = F, col.names = T, qmethod = c("escape", "double")) -write(paste((Sys.time() - now)/60, "min used for contrast calc"), file="/home-link/iivow01/git/rnadeseq/error/times", append=T) - ``` @@ -1675,7 +1653,6 @@ config(pg, modeBarButtonsToAdd = list(svg_exp)) %>% layout(margin = list(l=80, b ``` ```{r box_plots_noBatch, echo=F, message=F} -now <- Sys.time() # Decide which object to use for boxplots without batch correction if (run_rlog) { box_object <- rld_noncorrected @@ -1827,8 +1804,6 @@ if (!inherits(df_gene, 'try-error')){ names = rownames(df_gene) gene = TRUE } -write(paste((Sys.time() - now)/60, "min used for boxplot stuff"), file="/home-link/iivow01/git/rnadeseq/error/times", append=T) -nownow <- Sys.time() ``` ```{r genes_block, echo=FALSE, results='asis', eval=gene} @@ -1841,7 +1816,7 @@ cat("Plots for the [genes of interest](./differential_gene_expression/metadata/g ```{r load_PA_libs, eval=params$pathway_analysis, echo=FALSE, message=FALSE, warning=FALSE, results = 'hide'} -now <- Sys.time() + # Load libs invisibly invisible(lapply(c( "gprofiler2", @@ -1870,7 +1845,7 @@ species_library_installed <- get(params$species_library) if (isProvided(params$datasources)) { datasources <- unlist(strsplit(params$datasources, ',')) } else { - datasources <- c("KEGG", "REAC") + datasources <- c() } # Default to 1 for the nb of genes DE in a pathway @@ -1991,7 +1966,6 @@ for (file in contrast_files){ # Define list of Ensemble IDs (q) to run the pathway analysis q = as.character(DE_genes$Ensembl_ID) - write(paste((Sys.time() - nownow)/60, "min used for weird slow stuff"), file="/home-link/iivow01/git/rnadeseq/error/times", append=T) # gost query if (isProvided(params$set_background)) { @@ -2127,7 +2101,6 @@ for (file in contrast_files){ # Plotting heatmaps and KEGG pathways for all pathways print("Plotting heatmaps...") if (nrow(df) <= 100 & nrow(df) > 0) { - capture.output(df, file="/home-link/iivow01/git/rnadeseq/error/df.tsv", sep="\t", quote=F) conditions <- grepl("Condition", colnames(metadata)) metadata_save <- as.data.frame(metadata[,conditions]) metadata_name <- metadata[,c("QBiC.Code", "Secondary.Name")] @@ -2135,15 +2108,8 @@ for (file in contrast_files){ for (i in c(1:nrow(df))){ pathway <- df[i,] - capture.output(pathway, file="/home-link/iivow01/git/rnadeseq/error/pathway.tsv", sep="\t", quote=F) - gene_list <- unlist(strsplit(pathway$intersection, ",")) - write(paste((Sys.time() - now)/60, "min used for PA before mat stuff"), file="/home-link/iivow01/git/rnadeseq/error/times", append=T) - - capture.output(gene_list, file="/home-link/iivow01/git/rnadeseq/error/gene_list") - mat <- norm_counts[gene_list, ] - write.table(mat, file="/home-link/iivow01/git/rnadeseq/error/mat.tsv", sep="\t", quote=F) - write.table(norm_counts, file="/home-link/iivow01/git/rnadeseq/error/norm_countsss.tsv", sep="\t", quote=F) + mat <- norm_counts[which(norm_counts$Ensembl_ID %in% gene_list), ] rownames(mat) <- mat$gene_name mat$gene_name <- NULL mat <- data.matrix(mat) @@ -2250,8 +2216,7 @@ if (isProvided(params$path_genelist)){ gene_list <- na.omit(gene_list) rownames(norm_counts) <- norm_counts$Ensembl_ID - mat <- norm_counts[gene_list, ] - + mat <- norm_counts[which(norm_counts$Ensembl_ID %in% gene_list), ] rownames(mat) <- mat$gene_name mat$gene_name <- NULL mat$Ensembl_ID <- NULL @@ -2414,8 +2379,6 @@ You can find these heatmaps inside the [pathway_analysis](./pathway_analysis) fo and the subfolder: `pathway_heatmaps`. \ ")) -write(paste((Sys.time() - now)/60, "min used for all PA stuff"), file="/home-link/iivow01/git/rnadeseq/error/times", append=T) - ``` @@ -2619,4 +2582,4 @@ sessioninfo::session_info()%>% [^6]: https://github.com/nf-core/smrnaseq [^7]: https://github.com/OpenGene/fastp -[^8]: https://github.com/friedlanderlab/mirtrace \ No newline at end of file +[^8]: https://github.com/friedlanderlab/mirtrace diff --git a/bin/Execute_report.R b/bin/Execute_report.R index 5b9160d1..cd95b52e 100755 --- a/bin/Execute_report.R +++ b/bin/Execute_report.R @@ -34,6 +34,7 @@ option_list = list( make_option("--set_background", action="store_true", default=TRUE, help="Whether to use a background list for pathway analysis; if true, will only consider expressed genes (i.e. mean counts > 0) for PA."), make_option("--custom_background", type="character", default=NULL, help="Path to a custom background list TXT for pathway analysis; if provided, will only consider genes in that list for PA."), make_option(c("-w", "--min_DEG_pathway"), type="integer", default=NULL, help="min. number of genes DE in a pathway for this pathway to be considered enriched.", metavar="integer"), + make_option("--datasources", type="character", default=NULL, help="Which datasources to use for pathway analysis.", metavar="character"), make_option(c("-s", "--proj_summary"), type="character", default=NULL, help="Project summary file", metavar="character"), make_option(c("--path_quote"), type="character", default=NULL, help="Path to the quote PDF", metavar="character"), @@ -82,6 +83,7 @@ rmarkdown::render(opt$report, output_file = opt$output, knit_root_dir = wd, outp custom_background = opt$custom_background, keytype = opt$keytype, min_DEG_pathway = opt$min_DEG_pathway, + datasources = opt$datasources, path_proj_summary = opt$proj_summary, path_quote = opt$path_quote, diff --git a/conf/test.config b/conf/test.config index 05a27ad8..5280f668 100644 --- a/conf/test.config +++ b/conf/test.config @@ -28,6 +28,7 @@ params { software_versions = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/software_versions.csv' multiqc = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/MultiQC.zip' run_pathway_analysis = true + datasources = 'KEGG,REAC' genome = 'GRCm38' quote = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/offer_example.pdf' } diff --git a/conf/test_custom_gmt.config b/conf/test_custom_gmt.config index e3f84934..3ee2eab9 100644 --- a/conf/test_custom_gmt.config +++ b/conf/test_custom_gmt.config @@ -28,6 +28,7 @@ params { software_versions = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/software_versions.csv' multiqc = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/MultiQC.zip' run_pathway_analysis = true + datasources = 'KEGG,REAC' genome = 'GRCm38' custom_gmt = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/gprofiler_full_mmusculus.ENSG.gmt' } diff --git a/conf/test_smrnaseq.config b/conf/test_smrnaseq.config index 797fa332..6291a66d 100644 --- a/conf/test_smrnaseq.config +++ b/conf/test_smrnaseq.config @@ -24,6 +24,7 @@ params { genelist = 'testdata/smrnaseq/requested_genes.txt' input_type = 'smrnaseq' run_pathway_analysis = true + datasources = 'KEGG,REAC' genome = 'GRCh37' batch_effect = true set_background = false // This is solely to get some enriched pathways so that the github tests also check the pathway analysis output diff --git a/conf/test_star_rsem.config b/conf/test_star_rsem.config index 03c11cd5..9e8000ce 100644 --- a/conf/test_star_rsem.config +++ b/conf/test_star_rsem.config @@ -23,9 +23,10 @@ params { //report_options = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/report_options.yml' project_summary = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/summary.tsv' software_versions = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/software_versions_rsem.yml' - multiqc = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/MultiQC.zip' + //multiqc = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/MultiQC.zip' input_type = 'rsem' run_pathway_analysis = true + datasources = 'GO:CC' genome = 'GRCh37' // species_library = "org.Hs.eg.db" // organism = "hsapiens" diff --git a/conf/test_star_salmon.config b/conf/test_star_salmon.config index 4a896dfe..99a9e1ed 100644 --- a/conf/test_star_salmon.config +++ b/conf/test_star_salmon.config @@ -26,6 +26,7 @@ params { multiqc = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/MultiQC.zip' input_type = 'salmon' run_pathway_analysis = true + datasources = 'KEGG,REAC' // genome = 'GRCh37' species_library = "org.Hs.eg.db" organism = "hsapiens" diff --git a/docs/output.md b/docs/output.md index 99131726..6cc4e044 100644 --- a/docs/output.md +++ b/docs/output.md @@ -52,8 +52,10 @@ If pathway analysis was run, this directory contains the zipped pathway analysis - `*_gost_pathway_venn_diagram.pdf/png` - Venn diagrams showing the numbers of enriched pathways when using a background gene list vs when not using a bg list. -- `*_KEGG_pathway_enrichment_plot.pdf/png` - - Barplots showing the proportion of differentially expressed genes in the pathway. +- `enrichment_plots/*_pathway_enrichment_plot.{pdf/png/svg}` + - Barplots showing the proportion of differentially expressed genes in the pathway for a certain pathway database. +- `gost_pathway_gostplot.{pdf/png/svg}` + - Manhattan plots displaying all enriched pathways. - `KEGG_pathways/` - Contains the KEGG pathways graphs with the log fold change of the differentially expressed genes. - `pathway_heatmaps` diff --git a/docs/usage.md b/docs/usage.md index 2d74b416..ba2dac7c 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -89,7 +89,7 @@ The `qbic-pipelines/rnadeseq` pipeline relies on the output from the `nf-core/rn The typical command for running the pipeline is as follows: ```bash -nextflow run qbic-pipelines/rnadeseq -r 2.0.1 -profile docker \ +nextflow run qbic-pipelines/rnadeseq -r 2.2 -profile docker \ --gene_counts 'merged_gene_counts.txt' \ --input 'QXXXX_sample_preparations.tsv' \ --model 'linear_model.txt' \ @@ -157,7 +157,7 @@ curl https://codeload.github.com/qbic-pipelines/rnadeseq/tar.gz/master | tar -xz Afterwards, you should be able to also run test_star_rsem and test_star_salmon from qbic-pipelines/rnadeseq without manually cloning, e.g.: ```bash -nextflow run qbic-pipelines/rnadeseq -r 2.0.1 -profile docker,test_star_salmon +nextflow run qbic-pipelines/rnadeseq -r 2.2 -profile docker,test_star_salmon ``` ### Reproducibility diff --git a/modules/local/report.nf b/modules/local/report.nf index 167fceb5..565aa66e 100644 --- a/modules/local/report.nf +++ b/modules/local/report.nf @@ -37,6 +37,7 @@ process REPORT { def custom_gmt_opt = custom_gmt.name != 'NO_FILE3' ? "--custom_gmt $custom_gmt" : '' def set_background_opt = params.set_background ? "--set_background TRUE" : "--set_background FALSE" def custom_background_opt = custom_background.name != 'NO_FILE7' ? "--custom_background $custom_background" : '' + def datasources_opt = params.datasources ? "--datasources $params.datasources" : '' def quote_opt = params.quote != 'NO_FILE5' ? "--path_quote $params.quote" : '' def software_versions_opt = params.software_versions != 'NO_FILE6' ? "--software_versions $params.software_versions" : '' @@ -47,7 +48,7 @@ process REPORT { if [ "$multiqc" != "NO_FILE4" ]; then unzip $multiqc mkdir QC - mv MultiQC/multiqc_plots/ MultiQC/multiqc_data/ MultiQC/multiqc_report.html QC/ + mv MultiQC/multiqc_plots/ MultiQC/multiqc_data/ MultiQC/multiqc_report.html QC/ || mv multiqc/*/multiqc_plots/ multiqc/*/multiqc_data/ multiqc/*/multiqc_report.html QC/ || mv multiqc_plots/ multiqc_data/ multiqc_report.html QC/ fi Execute_report.R \ --report '$baseDir/assets/RNAseq_report.Rmd' \ @@ -76,6 +77,7 @@ process REPORT { --species_library $params.species_library \ --keytype $params.keytype \ --min_DEG_pathway $params.min_DEG_pathway \ + $datasources_opt \ $quote_opt \ $software_versions_opt \ --proj_summary $proj_summary \ @@ -100,4 +102,3 @@ process REPORT { """ } - diff --git a/nextflow.config b/nextflow.config index 9018f037..d476cd80 100644 --- a/nextflow.config +++ b/nextflow.config @@ -39,6 +39,7 @@ params { custom_gmt = 'NO_FILE3' set_background = true custom_background = 'NO_FILE7' + datasources = null // Additional args for the report project_summary = null @@ -252,7 +253,7 @@ manifest { description = """Differential gene expression analysis and pathway analysis of RNAseq data""" mainScript = 'main.nf' nextflowVersion = '!>=23.04.0' - version = '2.2' + version = 'dev' doi = '' } diff --git a/nextflow_schema.json b/nextflow_schema.json index adddbf59..d41f8c26 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -125,7 +125,7 @@ }, "custom_gmt": { "type": "string", - "description": "Path to custom GMT file for gost query, this allows e.g. to query older versions of databases for pathway analysis, not necessary if --skip_pathway_analysis = true." + "description": "Path to custom GMT file for gost query, this allows e.g. to query older versions of databases for pathway analysis, not necessary if --skip_pathway_analysis = true. If --datasources is set, the GMT file will be filtered for these datasources." }, "set_background": { "type": "boolean", @@ -140,6 +140,10 @@ "type": "integer", "default": 1, "description": "Integer indicating how many genes in a pathway must be differentially expressed to be considered as enriched, and report these pathways in tables and the final report. The default value is 1." + }, + "datasources": { + "type": "string", + "description": "Which datasources to use for pathway analysis, comma-separated string like 'KEGG,REAC'. See param 'sources' on https://rdrr.io/cran/gprofiler2/man/gost.html for a list of available sources. If not set, will use all sources. If set while a --custom_gmt is provided, will filter the GMT for these datasources (will not filter for the GO subtypes like GO:BP, just for GO)." } } }, diff --git a/tests/test.yml b/tests/test.yml old mode 100644 new mode 100755 index 56ff217d..585f4a04 --- a/tests/test.yml +++ b/tests/test.yml @@ -60,35 +60,36 @@ - path: results_test/differential_gene_expression/plots/PCA_plot.pdf - path: results_test/differential_gene_expression/plots/PCA_plot.png - path: results_test/differential_gene_expression/plots/PCA_plot.svg - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_venn_diagram.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_venn_diagram.png - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_venn_diagram.svg - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_pathway_enrichment_results.tsv - md5sum: be0f292a205f2ecd453ee27ecdc62615 - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_results.tsv - md5sum: f0e6d8117e3ff05ddabc3abb98f514b7 - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_results.tsv - md5sum: 3d7616fd0f6cb3aaf9f08252b2bcd223 - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_plot.png + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_venn_diagram.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_venn_diagram.png + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_venn_diagram.svg + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_gostplot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_gostplot.png + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_gostplot.svg + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/pathway_enrichment_results.tsv + md5sum: 21b907d52008f66f9a85190400fd211e + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathway_enrichment_results.tsv + md5sum: 92115e662e0e2489ad05d28f3981fa3a + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/REAC_pathway_enrichment_results.tsv + md5sum: 9077c63139668fd01d3f4086e6acb7bc + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/enrichment_plots/KEGG_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/enrichment_plots/KEGG_pathway_enrichment_plot.png + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/enrichment_plots/REAC_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/enrichment_plots/REAC_pathway_enrichment_plot.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04360.DE_contrast_condition_genotype_WT_vs_KO.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04360.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04360.xml - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04610.DE_contrast_condition_genotype_WT_vs_KO.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04610.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04610.xml - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_pathway_enrichment_results.tsv - md5sum: 0bd6dfd32bad6f5292f6cde91f019844 - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_results.tsv - md5sum: 452ca5dd8d5f190648988502f85d3ef1 - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.png - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.svg - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_plot.png + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/pathway_enrichment_results.tsv + md5sum: bdc0167b8b33a0da44e34768eead8376 + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/REAC_pathway_enrichment_results.tsv + md5sum: 6594fdb8c0eb591fa131982a8be00877 + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/gost_pathway_venn_diagram.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/gost_pathway_venn_diagram.png + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/gost_pathway_venn_diagram.svg + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/enrichment_plots/REAC_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/enrichment_plots/REAC_pathway_enrichment_plot.png - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf - path: results_test/RNAseq_report.html diff --git a/tests/test_batcheffect.yml b/tests/test_batcheffect.yml old mode 100644 new mode 100755 diff --git a/tests/test_contrast_list.yml b/tests/test_contrast_list.yml old mode 100644 new mode 100755 diff --git a/tests/test_contrast_matrix.yml b/tests/test_contrast_matrix.yml old mode 100644 new mode 100755 diff --git a/tests/test_custom_gmt.yml b/tests/test_custom_gmt.yml old mode 100644 new mode 100755 index 589ccf13..ea161a15 --- a/tests/test_custom_gmt.yml +++ b/tests/test_custom_gmt.yml @@ -60,11 +60,15 @@ - path: results_test/differential_gene_expression/plots/PCA_plot.pdf - path: results_test/differential_gene_expression/plots/PCA_plot.png - path: results_test/differential_gene_expression/plots/PCA_plot.svg - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_pathway_enrichment_results.tsv + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_gostplot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_gostplot.png + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_gostplot.svg + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_venn_diagram.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_venn_diagram.png + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_venn_diagram.svg + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/pathway_enrichment_results.tsv md5sum: eae154d9c7a526cc8c3fcd1a586fa0ed - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_pathway_enrichment_results.tsv + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/pathway_enrichment_results.tsv md5sum: bd2cbbf85c3e241743e60b9b8263a31c - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf - path: results_test/RNAseq_report.html diff --git a/tests/test_full.yml b/tests/test_full.yml old mode 100644 new mode 100755 index c4477cbf..beed5c7d --- a/tests/test_full.yml +++ b/tests/test_full.yml @@ -60,26 +60,26 @@ - path: results_test/differential_gene_expression/plots/PCA_plot.pdf - path: results_test/differential_gene_expression/plots/PCA_plot.png - path: results_test/differential_gene_expression/plots/PCA_plot.svg - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO_gost_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_pathway_enrichment_results.tsv - md5sum: be0f292a205f2ecd453ee27ecdc62615 - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_results.tsv - md5sum: f0e6d8117e3ff05ddabc3abb98f514b7 - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_results.tsv - md5sum: 3d7616fd0f6cb3aaf9f08252b2bcd223 - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_KEGG_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/DE_contrast_condition_genotype_WT_vs_KO_REAC_pathway_enrichment_plot.png + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_gostplot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_gostplot.png + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/pathway_enrichment_results.tsv + md5sum: 21b907d52008f66f9a85190400fd211e + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathway_enrichment_results.tsv + md5sum: 92115e662e0e2489ad05d28f3981fa3a + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/REAC_pathway_enrichment_results.tsv + md5sum: 9077c63139668fd01d3f4086e6acb7bc + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/enrichment_plots/KEGG_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/enrichment_plots/KEGG_pathway_enrichment_plot.png + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/enrichment_plots/REAC_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/enrichment_plots/REAC_pathway_enrichment_plot.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04360.DE_contrast_condition_genotype_WT_vs_KO.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04360.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04360.xml - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_pathway_enrichment_results.tsv - md5sum: 0bd6dfd32bad6f5292f6cde91f019844 - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_results.tsv - md5sum: 452ca5dd8d5f190648988502f85d3ef1 - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/DE_contrast_condition_treatment_Treated_vs_Control_REAC_pathway_enrichment_plot.png + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/pathway_enrichment_results.tsv + md5sum: bdc0167b8b33a0da44e34768eead8376 + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/REAC_pathway_enrichment_results.tsv + md5sum: 6594fdb8c0eb591fa131982a8be00877 + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/enrichment_plots/REAC_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/enrichment_plots/REAC_pathway_enrichment_plot.png - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf - path: results_test/RNAseq_report.html diff --git a/tests/test_no_multiqc.yml b/tests/test_no_multiqc.yml old mode 100644 new mode 100755 diff --git a/tests/test_relevel.yml b/tests/test_relevel.yml old mode 100644 new mode 100755 diff --git a/tests/test_smrnaseq.yml b/tests/test_smrnaseq.yml old mode 100644 new mode 100755 index 34ff104b..39676c01 --- a/tests/test_smrnaseq.yml +++ b/tests/test_smrnaseq.yml @@ -72,18 +72,18 @@ - path: results_test/differential_gene_expression/plots/PCA_plot_with_labels.pdf - path: results_test/differential_gene_expression/plots/PCA_plot_with_labels.png - path: results_test/differential_gene_expression/plots/PCA_plot_with_labels.svg - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/DE_contrast_condition_treatment_clone9_vs_clone1_KEGG_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/DE_contrast_condition_treatment_clone9_vs_clone1_KEGG_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/DE_contrast_condition_treatment_clone9_vs_clone1_KEGG_pathway_enrichment_plot.svg - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/DE_contrast_condition_treatment_clone9_vs_clone1_KEGG_pathway_enrichment_results.tsv - md5sum: 2637f5f6f4e74e7723280449e3ac647b - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/DE_contrast_condition_treatment_clone9_vs_clone1_pathway_enrichment_results.tsv - md5sum: a091e2ae9a678599d9e994082907d646 - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/DE_contrast_condition_treatment_control_vs_clone1_KEGG_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/DE_contrast_condition_treatment_control_vs_clone1_KEGG_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/DE_contrast_condition_treatment_control_vs_clone1_KEGG_pathway_enrichment_plot.svg - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/DE_contrast_condition_treatment_control_vs_clone1_KEGG_pathway_enrichment_results.tsv - md5sum: 497cbe026880c9ea78c5591c7c4d07f7 - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/DE_contrast_condition_treatment_control_vs_clone1_pathway_enrichment_results.tsv - md5sum: 63a2adf7c4a971dc7443503cf968e455 + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/enrichment_plots/KEGG_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/enrichment_plots/KEGG_pathway_enrichment_plot.png + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/enrichment_plots/KEGG_pathway_enrichment_plot.svg + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/KEGG_pathway_enrichment_results.tsv + md5sum: 326f3495ca42dbc527484cef3ee0e58d + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/pathway_enrichment_results.tsv + md5sum: b77e2839aa95ce3a9562d03a13629eb4 + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/enrichment_plots/KEGG_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/enrichment_plots/KEGG_pathway_enrichment_plot.png + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/enrichment_plots/KEGG_pathway_enrichment_plot.svg + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/KEGG_pathway_enrichment_results.tsv + md5sum: 64b3a3716b779900e99f56688c4b27e8 + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/pathway_enrichment_results.tsv + md5sum: 05e83209e36face6ead8b066155fdbe5 - path: results_test/RNAseq_report.html diff --git a/tests/test_star_rsem.yml b/tests/test_star_rsem.yml old mode 100644 new mode 100755 index c9cbdd49..d0d4b11c --- a/tests/test_star_rsem.yml +++ b/tests/test_star_rsem.yml @@ -75,38 +75,43 @@ - path: results_test/differential_gene_expression/plots/PCA_plot.pdf - path: results_test/differential_gene_expression/plots/PCA_plot.png - path: results_test/differential_gene_expression/plots/PCA_plot.svg - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_KEGG_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_KEGG_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_KEGG_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_KEGG_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_KEGG_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_KEGG_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_REAC_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_REAC_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_REAC_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_REAC_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_REAC_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_REAC_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_pathway_enrichment_results.tsv - md5sum: b1c0a1374c963fff71083a9cdefe8e9b - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_pathway_enrichment_results.tsv - md5sum: 631e89b4de9042936fa2652bde036e7d - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_pathway_enrichment_results.tsv - md5sum: 80123c4ba2c97eaa0217816f41122855 - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_KEGG_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_KEGG_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_KEGG_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_KEGG_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_KEGG_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_KEGG_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_REAC_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_REAC_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_REAC_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_REAC_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_REAC_pathway_enrichment_plot.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/DE_contrast_condition_cellline_MCF7_vs_GM12878_REAC_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_KEGG_pathway_enrichment_results.tsv - md5sum: 276e871c81e85217bbd7eef951676084 - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_REAC_pathway_enrichment_results.tsv - md5sum: 9e6847fe5d631f09305dfa00dd3472d1 - - path: results_test/RNAseq_report.html + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.png + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.svg + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/gost_pathway_gostplot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/gost_pathway_gostplot.png + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/gost_pathway_gostplot.svg + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/gost_pathway_venn_diagram.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/gost_pathway_venn_diagram.png + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/gost_pathway_venn_diagram.svg + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/pathway_enrichment_results.tsv + md5sum: 9eef4eab3da88b4b7e465f982be72acb + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/GO.CC_pathway_enrichment_results.tsv + md5sum: 2bbbf2de30f9da36e4c5b2df7a4ef5e7 + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.png + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.svg + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/gost_pathway_gostplot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/gost_pathway_gostplot.png + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/gost_pathway_gostplot.svg + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/gost_pathway_venn_diagram.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/gost_pathway_venn_diagram.png + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/gost_pathway_venn_diagram.svg + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/pathway_enrichment_results.tsv + md5sum: cbda28eeba947c53847ea28fdaa0cebd + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/GO.CC_pathway_enrichment_results.tsv + md5sum: 6b79ce64567f94ad3fc492e1a56fce58 + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.png + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.svg + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/gost_pathway_gostplot.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/gost_pathway_gostplot.png + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/gost_pathway_gostplot.svg + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/gost_pathway_venn_diagram.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/gost_pathway_venn_diagram.png + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/gost_pathway_venn_diagram.svg + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/pathway_enrichment_results.tsv + md5sum: 68f919b58d7fe5cc1b9a38665abccf78 + - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf + - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.svg + - path: results_test/pathway_analysis/metadata/gprofiler_full_hsapiens.ENSG.gmt diff --git a/tests/test_star_salmon.yml b/tests/test_star_salmon.yml old mode 100644 new mode 100755 index dbe49c9a..9d693acb --- a/tests/test_star_salmon.yml +++ b/tests/test_star_salmon.yml @@ -69,8 +69,8 @@ - path: results_test/differential_gene_expression/plots/PCA_plot.pdf - path: results_test/differential_gene_expression/plots/PCA_plot.png - path: results_test/differential_gene_expression/plots/PCA_plot.svg - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/DE_contrast_condition_cellline_H1_vs_GM12878_pathway_enrichment_results.tsv - md5sum: cd967cc355045d9dc8c0bb6ffcf40b61 - - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/DE_contrast_condition_cellline_K562_vs_GM12878_pathway_enrichment_results.tsv - md5sum: 68b329da9893e34099c7d8ad5cb9c940 + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/pathway_enrichment_results.tsv + md5sum: 30af65ef53ef7e389e116c31d31bd1d6 + - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/pathway_enrichment_results.tsv + md5sum: 4a3e61c2b4fea94c327acc63c83c0a4a - path: results_test/RNAseq_report.html diff --git a/tests/test_use_vst.yml b/tests/test_use_vst.yml old mode 100644 new mode 100755 From 7b24e9cad70e7c8a91ef54cf0674a713f2ccc263 Mon Sep 17 00:00:00 2001 From: WackerO Date: Wed, 24 Apr 2024 13:49:42 +0200 Subject: [PATCH 29/73] Added multiqc.zips for the new salmon/rsem output --- assets/RNAseq_report.Rmd | 112 ++++++++++++++++++++------ conf/test_star_rsem.config | 5 +- conf/test_star_salmon.config | 2 +- testdata/QDESQ/new_rsem_multiqc.zip | Bin 0 -> 753582 bytes testdata/QDESQ/new_salmon_multiqc.zip | Bin 0 -> 753954 bytes 5 files changed, 89 insertions(+), 30 deletions(-) create mode 100644 testdata/QDESQ/new_rsem_multiqc.zip create mode 100644 testdata/QDESQ/new_salmon_multiqc.zip diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index 921dbf1b..a2ac62d1 100755 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -896,6 +896,17 @@ if (file.exists(paste0(wd,"/QC/multiqc_data/multiqc_general_stats.txt"))) { is.num <- sapply(mqc_stats, is.numeric) mqc_stats[is.num] <- lapply(mqc_stats[is.num], round, 2) +# The following gets rid of multiQC entries that are spread across two rows, one ending with -fw (see https://github.com/qbic-pipelines/rnadeseq/issues/114) +sum_NA <- function(x) {if (all(is.na(x))) x[NA_integer_] else sum(x, na.rm = TRUE)} # Function to sum across rows if available, else NA +mqc_stats$Sample <- gsub("-fw", "", mqc_stats$Sample) # Remove -fw to produce duplicate rows for summarising +mqc_stats <- mqc_stats %>% group_by(Sample) %>% summarise_all(sum_NA) + +# In newer RNASeq versions (after 1.4.2?), duplicate rows might not end with -fw, but with _1 and _2 instead; deal with those in the following lines +avg_NA <- function(x) {if (all(is.na(x))) x[NA_integer_] else mean(x, na.rm = TRUE)} # Function to average across rows if available, else NA +mqc_stats$Sample <- sub("_[0-9]+$", "", mqc_stats$Sample) # Remove _1 etc. to produce duplicate rows for summarising +mqc_stats <- mqc_stats %>% group_by(Sample) %>% summarise_all(avg_NA) + + if (mqc_version == 'old_mqc') { columns <- c( "Sample", @@ -910,37 +921,77 @@ if (mqc_version == 'old_mqc') { columns <- c( "Sample", "raw_total_sequences", - "reads_duplicated_percent", - "reads_mapped_percent" + "reads_mapped_percent", + "reads_duplicated_percent" ) + + # For the new version, some more tables need to be read + if (params$input_type == "salmon" && file.exists(paste0(wd,"/QC/multiqc_data/multiqc_star.txt"))) { + star <- read.table(file = paste0(wd,"/QC/multiqc_data/multiqc_star.txt"), header=TRUE, sep="\t", check.names = F)[,c("Sample", "uniquely_mapped_percent")] + } else if (params$input_type == "rsem" && file.exists(paste0(wd,"/QC/multiqc_data/multiqc_rsem.txt"))) { + star <- read.table(file = paste0(wd,"/QC/multiqc_data/multiqc_rsem.txt"), header=TRUE, sep="\t", check.names = F)[,c("Sample", "Alignable", "Unique")] + star$uniquely_mapped_percent <- star$Unique/star$Alignable + star$Unique <- NULL + star$Alignable <- NULL + } else { + star <- NULL + } + if (file.exists(paste0(wd,"/QC/multiqc_data/multiqc_cutadapt.txt"))) { + cutadapt <- read.table(file = paste0(wd,"/QC/multiqc_data/multiqc_cutadapt.txt"), header=TRUE, sep="\t", check.names = F)[,c("Sample", "percent_trimmed")] + cutadapt$Sample <- sub("_[0-9]+$", "", cutadapt$Sample) # Remove _1 etc. to produce duplicate rows for summarising + cutadapt <- cutadapt %>% group_by(Sample) %>% summarise_all(sum_NA) + } else { + cutadapt <- NULL + } + if (file.exists(paste0(wd,"/QC/multiqc_data/multiqc_rseqc_read_distribution.txt"))) { + rseqc <- read.table(file = paste0(wd,"/QC/multiqc_data/multiqc_rseqc_read_distribution.txt"), header=TRUE, sep="\t", check.names = F)[,c("Sample", "total_tags", "total_assigned_tags")] + } else { + rseqc <- NULL + } + if (file.exists(paste0(wd,"/QC/multiqc_data/multiqc_fastqc.txt"))) { + fastqc_untrimmed <- read.table(file = paste0(wd,"/QC/multiqc_data/multiqc_fastqc.txt"), header=TRUE, sep="\t", check.names = F)[,c("Sample", "%GC")] + colnames(fastqc_untrimmed)[colnames(fastqc_untrimmed) == "%GC"] <- "GC_untrimmed" + fastqc_untrimmed$Sample <- sub("_[0-9]+$", "", fastqc_untrimmed$Sample) # Remove _1 etc. to produce duplicate rows for summarising + fastqc_untrimmed <- fastqc_untrimmed %>% group_by(Sample) %>% summarise_all(sum_NA) + } else { + fastqc_untrimmed <- NULL + } + if (file.exists(paste0(wd,"/QC/multiqc_data/multiqc_fastqc_1.txt"))) { + fastqc_trimmed <- read.table(file = paste0(wd,"/QC/multiqc_data/multiqc_fastqc_1.txt"), header=TRUE, sep="\t", check.names = F)[,c("Sample", "%GC")] + colnames(fastqc_trimmed)[colnames(fastqc_trimmed) == "%GC"] <- "GC_trimmed" + fastqc_trimmed$Sample <- sub("_[0-9]+$", "", fastqc_trimmed$Sample) # Remove _1 etc. to produce duplicate rows for summarising + fastqc_trimmed <- fastqc_trimmed %>% group_by(Sample) %>% summarise_all(sum_NA) + } else { + fastqc_trimmed <- NULL + } } -mqc_stats_selected <- mqc_stats[,columns] -# The following gets rid of multiQC entries that are spread across two rows, one ending with -fw (see https://github.com/qbic-pipelines/rnadeseq/issues/114) -sum_NA <- function(x) {if (all(is.na(x))) x[NA_integer_] else sum(x, na.rm = TRUE)} # Function to sum across rows if available, else NA -mqc_stats_selected$Sample <- gsub("-fw", "", mqc_stats_selected$Sample) # Remove -fw to produce duplicate rows for summarising -mqc_stats_selected <- mqc_stats_selected %>% group_by(Sample) %>% summarise_all(sum_NA) +table_complete <- mqc_stats[,columns] -# In newer RNASeq versions (after 1.4.2?), duplicate rows might not end with -fw, but with _1 and _2 instead; deal with those in the following lines -avg_NA <- function(x) {if (all(is.na(x))) x[NA_integer_] else mean(x, na.rm = TRUE)} # Function to average across rows if available, else NA -mqc_stats_selected$Sample <- sub("_[0-9]+$", "", mqc_stats_selected$Sample) # Remove _1 etc. to produce duplicate rows for summarising -mqc_stats_selected <- mqc_stats_selected %>% group_by(Sample) %>% summarise_all(avg_NA) +if (mqc_version == 'new_mqc') { + + # Merge the additional tables into the stats + if (!is.null(cutadapt)) table_complete <- merge(table_complete,cutadapt,by="Sample") + if (!is.null(rseqc)) table_complete <- merge(table_complete,rseqc,by="Sample") + if (!is.null(star)) table_complete <- merge(table_complete,star,by="Sample") + if (!is.null(fastqc_untrimmed)) table_complete <- merge(table_complete,fastqc_untrimmed,by="Sample") + if (!is.null(fastqc_trimmed)) table_complete <- merge(table_complete,fastqc_trimmed,by="Sample") +} -mqc_stats_selected$Sample <- substr(mqc_stats_selected$Sample, 1, 10) -n_rows = nrow(mqc_stats_selected) +# Reduce to QBiC code +table_complete$Sample <- substr(table_complete$Sample, 1, 10) metadata <- read.table((paste0(wd, "/differential_gene_expression/metadata/metadata.tsv")), header=TRUE, sep="\t") -metadata = metadata[,c("QBiC.Code")] +metadata <- metadata[,c("QBiC.Code")] #the following makes metadata a df again (in R, when extracting only 1 column from a df, it becomes a list) metadata <- data.frame(matrix(unlist(metadata))) colnames(metadata) <- c("Sample") - -table_complete = merge(metadata,mqc_stats_selected,by="Sample") - +table_complete <- merge(metadata,table_complete,by="Sample") if (mqc_version == 'old_mqc') { - colnames = c("Sample", "Number of reads (M)", "Duplicates (%)", "GC (%)", "Trimmed reads (%)", "Mapped reads (%)", "Assigned reads (%)") + colnames <- c("Sample", "Number of reads (M)", "Duplicates (%)", "GC (%)", "Trimmed reads (%)", "Mapped reads (%)", "Assigned reads (%)") colnames(table_complete) <- c("Sample", "ReadNumber", "DuplicateReadsIntercept", "GCcontent", "TrimmedReads", "MappedReads", "AssignedReads") + table_complete <- table_complete %>% mutate( Sample = Sample, @@ -958,16 +1009,25 @@ if (mqc_version == 'old_mqc') { color_bar("orange")(AssignedReads)) ) } else { - colnames = c("Sample", "Number of reads (M)", "Duplicates (%)", "Mapped reads (%)") - colnames(table_complete) <- c("Sample", "ReadNumber", "DuplicateReadsIntercept", "MappedReads") + colnames <- c("Sample", "Number of reads (M)", "Duplicates (%)", "Untrimmed GC (%)", "Trimmed GC (%)", "Trimmed reads (%)", "Mapped reads (%)", "Assigned reads (%)") + #colnames(table_complete) <- c("Sample", "ReadNumber", "DuplicateReadsIntercept") + table_complete <- table_complete %>% - mutate( + transmute( Sample = Sample, - ReadNumber = color_bar("lightblue")(round((ReadNumber/1000000),2)), - DuplicateReadsIntercept = ifelse(DuplicateReadsIntercept > 1, - cell_spec(DuplicateReadsIntercept, color="orange", bold=T), - cell_spec(DuplicateReadsIntercept, color="black")), - MappedReads = ifelse(MappedReads > 80, color_bar("lightblue")(MappedReads), color_bar("orange")(MappedReads)) + ReadNumber = color_bar("lightblue")(round((raw_total_sequences/1000000),2)), + DuplicateReadsIntercept = ifelse(reads_duplicated_percent > 1, + cell_spec(reads_duplicated_percent, color="orange", bold=T), + cell_spec(reads_duplicated_percent, color="black")), + GC_untrimmed = GC_untrimmed, + GC_trimmed = GC_trimmed, + TrimmedReads = percent_trimmed, + MappedReads = ifelse(uniquely_mapped_percent > 80, + color_bar("lightblue")(uniquely_mapped_percent), + color_bar("orange")(uniquely_mapped_percent)), + AssignedReads = ifelse(total_assigned_tags/total_tags > 60, + color_bar("lightblue")(total_assigned_tags/total_tags), + color_bar("orange")(total_assigned_tags/total_tags)) ) } diff --git a/conf/test_star_rsem.config b/conf/test_star_rsem.config index 54d63a30..840a5191 100755 --- a/conf/test_star_rsem.config +++ b/conf/test_star_rsem.config @@ -23,12 +23,11 @@ params { //report_options = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/report_options.yml' project_summary = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/summary.tsv' software_versions = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/software_versions_rsem.yml' - //multiqc = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/MultiQC.zip' + multiqc = 'testdata/QDESQ/new_rsem_multiqc.zip' input_type = 'rsem' run_pathway_analysis = true - datasources = 'GO:CC' + datasources = 'GO:CC,KEGG' genome = 'GRCh37' - datasources = 'GO:CC' // species_library = "org.Hs.eg.db" // organism = "hsapiens" // keytype = "ENSEMBL" diff --git a/conf/test_star_salmon.config b/conf/test_star_salmon.config index 99a9e1ed..708c532c 100755 --- a/conf/test_star_salmon.config +++ b/conf/test_star_salmon.config @@ -23,7 +23,7 @@ params { //report_options = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/report_options.yml' project_summary = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/summary.tsv' software_versions = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/software_versions_salmon.yml' - multiqc = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/MultiQC.zip' + multiqc = 'testdata/QDESQ/new_salmon_multiqc.zip' input_type = 'salmon' run_pathway_analysis = true datasources = 'KEGG,REAC' diff --git a/testdata/QDESQ/new_rsem_multiqc.zip 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zTS!>Q-<->B`Hx_uC+}~M$%kS@Z{s22E|F>wx&799~ zE3TB+BRVM9Ezrd&$j#Z+$<-@3B*^PtXowfs{~t z|D*r4A;0$3@jnFs0HS~y1B?CtUO<2^I3)Ow#sZi9i~9b9a~%0ANA{mMG=q}=iW3a+ z_=Ba?_%B$0Ht{AaEYzi;@+bgs_cZ`u`cJ05iP|Oh-{kZ=%kTGE{#!+R`MbLXIk|-e zxCMFnx%r1U{hs({hiyLVZ_WK*2+Dk|Y1DlN0ASMqfNQ@9GF1KRg8rWt`yVJ;p_bn% zR{#K}76AZ@UlhG|`rlLJ6yWk#Y+3<3!9q>}0H9_7z~mQEeQ|$9RPf)s!P(c#!~cJ{ zX1 Date: Wed, 24 Apr 2024 14:58:22 +0200 Subject: [PATCH 30/73] moved heatmap_genelist output to differential analysis results folder --- assets/RNAseq_report.Rmd | 10 +++++----- 1 file changed, 5 insertions(+), 5 deletions(-) diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index ae01a018..624e01b7 100755 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -2128,8 +2128,8 @@ for (file in contrast_files){ if (isProvided(params$path_genelist)){ genelist_path = params$path_genelist - genelist_heatmaps_dir <- "heatmap_gene_list" - dir.create(paste("pathway_analysis", genelist_heatmaps_dir, sep="/")) + genelist_heatmaps_dir <- "differential_gene_expression/plots/heatmap_gene_list" + dir.create(genelist_heatmaps_dir) print("Plotting heatmaps...") conditions <- grepl("Condition", colnames(metadata)) @@ -2158,13 +2158,13 @@ if (isProvided(params$path_genelist)){ mat <- data.matrix(mat) if (nrow(mat)>1){ - png(filename = paste0("pathway_analysis", "/", genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.png"), width = 100+ncol(mat)*50, height = 50+nrow(mat)*100) + png(filename = paste0(genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.png"), width = 100+ncol(mat)*50, height = 50+nrow(mat)*100) pheatmap(mat = mat, annotation_col = metadata_save, main = "Genelist-specific heatmap of normalized expression values for genes", scale = "row", cluster_cols = F, cluster_rows = T) dev.off() - pdf(paste0("pathway_analysis", "/", genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.pdf"), width = 10+ncol(mat)*0.05, height = 5+nrow(mat)*0.005) + pdf(paste0(genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.pdf"), width = 10+ncol(mat)*0.05, height = 5+nrow(mat)*0.005) pheatmap(mat = mat, annotation_col = metadata_save, main = "Genelist-specific heatmap of normalized expression values for genes", scale = "row", cluster_cols = F, cluster_rows = T) dev.off() - svg(paste0("pathway_analysis", "/", genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.svg"), width = 10+ncol(mat)*0.1, height = 5+nrow(mat)*0.005) + svg(paste0(genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.svg"), width = 10+ncol(mat)*0.1, height = 5+nrow(mat)*0.005) pheatmap(mat = mat, annotation_col = metadata_save, main = "Genelist-specific heatmap of normalized expression values for genes", scale = "row", cluster_cols = F, cluster_rows = T) dev.off() } From 582e71fa335e3e05823ced85d78ea971d6bc9c32 Mon Sep 17 00:00:00 2001 From: WackerO Date: Wed, 24 Apr 2024 15:25:28 +0200 Subject: [PATCH 31/73] removed ensembl_id col from pathway heatmaps --- assets/RNAseq_report.Rmd | 1 + 1 file changed, 1 insertion(+) diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index 624e01b7..0c23711b 100755 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -2047,6 +2047,7 @@ for (file in contrast_files){ mat <- norm_counts[gene_list, ] rownames(mat) <- mat$gene_name mat$gene_name <- NULL + mat$Ensembl_ID <- NULL mat <- data.matrix(mat) # Skip the heatmaps for CI tests because especially png() works only very unreliably for the heatmaps for some reason From ebbcf0440f85fde6d688c51a53b4daee5bd3cc5e Mon Sep 17 00:00:00 2001 From: WackerO Date: Thu, 25 Apr 2024 16:17:42 +0200 Subject: [PATCH 32/73] fixed plotma issue with empty plots, added clutser param for pathway heatmaps --- assets/RNAseq_report.Rmd | 21 +++++++++++---------- bin/Execute_report.R | 4 ++++ modules/local/report.nf | 5 +++++ nextflow.config | 2 ++ nextflow_schema.json | 10 ++++++++++ 5 files changed, 32 insertions(+), 10 deletions(-) diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index 0c23711b..b00bfdec 100755 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -44,6 +44,8 @@ params: set_background: '' custom_background: '' datasources: '' + heatmaps_cluster_rows: '' + heatmaps_cluster_cols: '' #Additional args for the report path_proj_summary: '' @@ -254,7 +256,6 @@ invisible( lapply(c( ``` ```{r create_outdirs, echo=FALSE, message=FALSE, warning=FALSE, results = 'hide'} - # create directories needed ifelse(!dir.exists("differential_gene_expression"), dir.create("differential_gene_expression"), FALSE) dir.create("differential_gene_expression/metadata") @@ -779,13 +780,13 @@ res=0 for (i in resultsNames(cds)[-1]) { res = results(cds,name = i) pdf(paste("differential_gene_expression/plots/further_diagnostics_plots/all_results_MA_plot_",i,".pdf",sep="")) - plotMA(res,ylim = c(-4, 4)) + DESeq2::plotMA(res,ylim = c(-4, 4)) dev.off() png(paste("differential_gene_expression/plots/further_diagnostics_plots/all_results_MA_plot_",i,".png",sep="")) - plotMA(res,ylim = c(-4, 4)) + DESeq2::plotMA(res,ylim = c(-4, 4)) dev.off() svg(paste("differential_gene_expression/plots/further_diagnostics_plots/all_results_MA_plot_",i,".svg",sep="")) - plotMA(res,ylim = c(-4, 4)) + DESeq2::plotMA(res,ylim = c(-4, 4)) dev.off() # multiple hyptothesis testing @@ -2058,7 +2059,7 @@ for (file in contrast_files){ tryCatch( { png(filename = paste0("pathway_analysis", "/",fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", make.names(pathway$term_id), ".png"), width = ifelse(nrow(mat)<=20, 200, 125)+ncol(mat)*40, height = 50+nrow(mat)*50) - pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T) + pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = params$heatmaps_cluster_cols, cluster_rows = params$heatmaps_cluster_rows) dev.off() }, error=function(e) { @@ -2068,7 +2069,7 @@ for (file in contrast_files){ tryCatch( { pdf(paste0("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", make.names(pathway$term_id), ".pdf"), width = ifelse(nrow(mat)<=20, 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005) - pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T) + pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = params$heatmaps_cluster_cols, cluster_rows = params$heatmaps_cluster_rows) dev.off() }, error=function(e) { @@ -2078,7 +2079,7 @@ for (file in contrast_files){ tryCatch( { svg(paste0("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", make.names(pathway$term_id), ".svg"), width = ifelse(nrow(mat)<=20, 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005) - pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = F, cluster_rows = T) + pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = params$heatmaps_cluster_cols, cluster_rows = params$heatmaps_cluster_rows) dev.off() }, error=function(e) { @@ -2160,13 +2161,13 @@ if (isProvided(params$path_genelist)){ if (nrow(mat)>1){ png(filename = paste0(genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.png"), width = 100+ncol(mat)*50, height = 50+nrow(mat)*100) - pheatmap(mat = mat, annotation_col = metadata_save, main = "Genelist-specific heatmap of normalized expression values for genes", scale = "row", cluster_cols = F, cluster_rows = T) + pheatmap(mat = mat, annotation_col = metadata_save, main = "Genelist-specific heatmap of normalized expression values for genes", scale = "row", cluster_cols = params$heatmaps_cluster_cols, cluster_rows = params$heatmaps_cluster_rows) dev.off() pdf(paste0(genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.pdf"), width = 10+ncol(mat)*0.05, height = 5+nrow(mat)*0.005) - pheatmap(mat = mat, annotation_col = metadata_save, main = "Genelist-specific heatmap of normalized expression values for genes", scale = "row", cluster_cols = F, cluster_rows = T) + pheatmap(mat = mat, annotation_col = metadata_save, main = "Genelist-specific heatmap of normalized expression values for genes", scale = "row", cluster_cols = params$heatmaps_cluster_cols, cluster_rows = params$heatmaps_cluster_rows) dev.off() svg(paste0(genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.svg"), width = 10+ncol(mat)*0.1, height = 5+nrow(mat)*0.005) - pheatmap(mat = mat, annotation_col = metadata_save, main = "Genelist-specific heatmap of normalized expression values for genes", scale = "row", cluster_cols = F, cluster_rows = T) + pheatmap(mat = mat, annotation_col = metadata_save, main = "Genelist-specific heatmap of normalized expression values for genes", scale = "row", cluster_cols = params$heatmaps_cluster_cols, cluster_rows = params$heatmaps_cluster_rows) dev.off() } } diff --git a/bin/Execute_report.R b/bin/Execute_report.R index cd95b52e..574fbc1e 100755 --- a/bin/Execute_report.R +++ b/bin/Execute_report.R @@ -35,6 +35,8 @@ option_list = list( make_option("--custom_background", type="character", default=NULL, help="Path to a custom background list TXT for pathway analysis; if provided, will only consider genes in that list for PA."), make_option(c("-w", "--min_DEG_pathway"), type="integer", default=NULL, help="min. number of genes DE in a pathway for this pathway to be considered enriched.", metavar="integer"), make_option("--datasources", type="character", default=NULL, help="Which datasources to use for pathway analysis.", metavar="character"), + make_option("--heatmaps_cluster_rows", action="store_true", default=FALSE, help="Whether to activate row clustering when generating heatmaps of gene expression in enriched pathways."), + make_option("--heatmaps_cluster_cols", action="store_true", default=FALSE, help="Whether to activate column clustering when generating heatmaps of gene expression in enriched pathways."), make_option(c("-s", "--proj_summary"), type="character", default=NULL, help="Project summary file", metavar="character"), make_option(c("--path_quote"), type="character", default=NULL, help="Path to the quote PDF", metavar="character"), @@ -84,6 +86,8 @@ rmarkdown::render(opt$report, output_file = opt$output, knit_root_dir = wd, outp keytype = opt$keytype, min_DEG_pathway = opt$min_DEG_pathway, datasources = opt$datasources, + heatmaps_cluster_rows = opt$heatmaps_cluster_rows, + heatmaps_cluster_cols = opt$heatmaps_cluster_cols, path_proj_summary = opt$proj_summary, path_quote = opt$path_quote, diff --git a/modules/local/report.nf b/modules/local/report.nf index a05ef3b7..5d53a709 100755 --- a/modules/local/report.nf +++ b/modules/local/report.nf @@ -38,6 +38,9 @@ process REPORT { def set_background_opt = params.set_background ? "--set_background TRUE" : "--set_background FALSE" def custom_background_opt = custom_background.name != 'NO_FILE7' ? "--custom_background $custom_background" : '' def datasources_opt = params.datasources ? "--datasources $params.datasources" : '' + def heatmaps_cluster_rows_opt = params.heatmaps_cluster_rows ? "--heatmaps_cluster_rows TRUE" : '' + def heatmaps_cluster_cols_opt = params.heatmaps_cluster_cols ? "--heatmaps_cluster_cols TRUE" : '' + def quote_opt = params.quote != 'NO_FILE5' ? "--path_quote $params.quote" : '' def software_versions_opt = params.software_versions != 'NO_FILE6' ? "--software_versions $params.software_versions" : '' @@ -78,6 +81,8 @@ process REPORT { --keytype $params.keytype \ --min_DEG_pathway $params.min_DEG_pathway \ $datasources_opt \ + $heatmaps_cluster_rows_opt \ + $heatmaps_cluster_cols_opt \ $quote_opt \ $software_versions_opt \ --proj_summary $proj_summary \ diff --git a/nextflow.config b/nextflow.config index 7397f9c5..d7581592 100755 --- a/nextflow.config +++ b/nextflow.config @@ -45,6 +45,8 @@ params { set_background = true custom_background = 'NO_FILE7' datasources = null + heatmaps_cluster_rows = true + heatmaps_cluster_cols = false // Additional args for the report project_summary = null diff --git a/nextflow_schema.json b/nextflow_schema.json index d41f8c26..b5639637 100755 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -144,6 +144,16 @@ "datasources": { "type": "string", "description": "Which datasources to use for pathway analysis, comma-separated string like 'KEGG,REAC'. See param 'sources' on https://rdrr.io/cran/gprofiler2/man/gost.html for a list of available sources. If not set, will use all sources. If set while a --custom_gmt is provided, will filter the GMT for these datasources (will not filter for the GO subtypes like GO:BP, just for GO)." + }, + "heatmaps_cluster_rows": { + "type": "boolean", + "default": true, + "description": "Whether to activate row clustering when generating heatmaps of gene expression in enriched pathways." + }, + "heatmaps_cluster_cols": { + "type": "boolean", + "default": false, + "description": "Whether to activate column clustering when generating heatmaps of gene expression in enriched pathways." } } }, From 5ac6a4f9b1711ed88493d8ce2aff0bc29b63659d Mon Sep 17 00:00:00 2001 From: WackerO Date: Thu, 25 Apr 2024 18:11:42 +0200 Subject: [PATCH 33/73] updated checksums --- tests/test.yml | 4 ++-- tests/test_smrnaseq.yml | 4 ++-- tests/test_star_rsem.yml | 10 +++++----- tests/test_star_salmon.yml | 4 ++-- 4 files changed, 11 insertions(+), 11 deletions(-) diff --git a/tests/test.yml b/tests/test.yml index 585f4a04..bc80982d 100755 --- a/tests/test.yml +++ b/tests/test.yml @@ -67,7 +67,7 @@ - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_gostplot.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_gostplot.svg - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/pathway_enrichment_results.tsv - md5sum: 21b907d52008f66f9a85190400fd211e + md5sum: 92736a4c802ebd4e644682308ecf46dc - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathway_enrichment_results.tsv md5sum: 92115e662e0e2489ad05d28f3981fa3a - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/REAC_pathway_enrichment_results.tsv @@ -83,7 +83,7 @@ - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04610.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/KEGG_pathways/mmu04610.xml - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/pathway_enrichment_results.tsv - md5sum: bdc0167b8b33a0da44e34768eead8376 + md5sum: 0a425c10e229bb775597be176c414705 - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/REAC_pathway_enrichment_results.tsv md5sum: 6594fdb8c0eb591fa131982a8be00877 - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/gost_pathway_venn_diagram.pdf diff --git a/tests/test_smrnaseq.yml b/tests/test_smrnaseq.yml index 39676c01..3cac7639 100755 --- a/tests/test_smrnaseq.yml +++ b/tests/test_smrnaseq.yml @@ -78,12 +78,12 @@ - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/KEGG_pathway_enrichment_results.tsv md5sum: 326f3495ca42dbc527484cef3ee0e58d - path: results_test/pathway_analysis/DE_contrast_condition_treatment_clone9_vs_clone1/pathway_enrichment_results.tsv - md5sum: b77e2839aa95ce3a9562d03a13629eb4 + md5sum: d63d4d96de3c49cc38c91cb863a7da06 - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/enrichment_plots/KEGG_pathway_enrichment_plot.pdf - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/enrichment_plots/KEGG_pathway_enrichment_plot.png - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/enrichment_plots/KEGG_pathway_enrichment_plot.svg - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/KEGG_pathway_enrichment_results.tsv md5sum: 64b3a3716b779900e99f56688c4b27e8 - path: results_test/pathway_analysis/DE_contrast_condition_treatment_control_vs_clone1/pathway_enrichment_results.tsv - md5sum: 05e83209e36face6ead8b066155fdbe5 + md5sum: e7a051d8623043a3849eae8900b99f71 - path: results_test/RNAseq_report.html diff --git a/tests/test_star_rsem.yml b/tests/test_star_rsem.yml index d0d4b11c..d14ab2d0 100755 --- a/tests/test_star_rsem.yml +++ b/tests/test_star_rsem.yml @@ -85,9 +85,9 @@ - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/gost_pathway_venn_diagram.png - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/gost_pathway_venn_diagram.svg - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/pathway_enrichment_results.tsv - md5sum: 9eef4eab3da88b4b7e465f982be72acb + md5sum: 57e56666c0d561659f5aaec478f04cee - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/GO.CC_pathway_enrichment_results.tsv - md5sum: 2bbbf2de30f9da36e4c5b2df7a4ef5e7 + md5sum: 098f12540b4cad2ea1c950af1fd304b6 - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.pdf - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.png - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.svg @@ -98,9 +98,9 @@ - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/gost_pathway_venn_diagram.png - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/gost_pathway_venn_diagram.svg - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/pathway_enrichment_results.tsv - md5sum: cbda28eeba947c53847ea28fdaa0cebd + md5sum: 73fdb835cede09c13f0ab2684b1cda73 - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/GO.CC_pathway_enrichment_results.tsv - md5sum: 6b79ce64567f94ad3fc492e1a56fce58 + md5sum: 2ebd84e4ed2029659294a09d49b62048 - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.pdf - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.png - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.svg @@ -111,7 +111,7 @@ - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/gost_pathway_venn_diagram.png - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/gost_pathway_venn_diagram.svg - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/pathway_enrichment_results.tsv - md5sum: 68f919b58d7fe5cc1b9a38665abccf78 + md5sum: b7f67bc86475b218965b173b753199b1 - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.svg - path: results_test/pathway_analysis/metadata/gprofiler_full_hsapiens.ENSG.gmt diff --git a/tests/test_star_salmon.yml b/tests/test_star_salmon.yml index 9d693acb..2b111164 100755 --- a/tests/test_star_salmon.yml +++ b/tests/test_star_salmon.yml @@ -70,7 +70,7 @@ - path: results_test/differential_gene_expression/plots/PCA_plot.png - path: results_test/differential_gene_expression/plots/PCA_plot.svg - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/pathway_enrichment_results.tsv - md5sum: 30af65ef53ef7e389e116c31d31bd1d6 + md5sum: 68b329da9893e34099c7d8ad5cb9c940 - path: results_test/pathway_analysis/DE_contrast_condition_cellline_K562_vs_GM12878/pathway_enrichment_results.tsv - md5sum: 4a3e61c2b4fea94c327acc63c83c0a4a + md5sum: 68b329da9893e34099c7d8ad5cb9c940 - path: results_test/RNAseq_report.html From acf798c97307597577cd0f8e78ba40b6aeba7683 Mon Sep 17 00:00:00 2001 From: WackerO Date: Sun, 28 Apr 2024 15:06:44 +0200 Subject: [PATCH 34/73] Bugfixing --- assets/RNAseq_report.Rmd | 35 +++++++++++++++++++++++------------ 1 file changed, 23 insertions(+), 12 deletions(-) diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index a2ac62d1..563eea31 100755 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -897,15 +897,14 @@ is.num <- sapply(mqc_stats, is.numeric) mqc_stats[is.num] <- lapply(mqc_stats[is.num], round, 2) # The following gets rid of multiQC entries that are spread across two rows, one ending with -fw (see https://github.com/qbic-pipelines/rnadeseq/issues/114) -sum_NA <- function(x) {if (all(is.na(x))) x[NA_integer_] else sum(x, na.rm = TRUE)} # Function to sum across rows if available, else NA mqc_stats$Sample <- gsub("-fw", "", mqc_stats$Sample) # Remove -fw to produce duplicate rows for summarising -mqc_stats <- mqc_stats %>% group_by(Sample) %>% summarise_all(sum_NA) # In newer RNASeq versions (after 1.4.2?), duplicate rows might not end with -fw, but with _1 and _2 instead; deal with those in the following lines -avg_NA <- function(x) {if (all(is.na(x))) x[NA_integer_] else mean(x, na.rm = TRUE)} # Function to average across rows if available, else NA mqc_stats$Sample <- sub("_[0-9]+$", "", mqc_stats$Sample) # Remove _1 etc. to produce duplicate rows for summarising -mqc_stats <- mqc_stats %>% group_by(Sample) %>% summarise_all(avg_NA) +# Now that the duplicate columns have been given the same names, we can group and collapse them with the appropriate functions +sum_NA <- function(x) {if (all(is.na(x))) x[NA_integer_] else sum(x, na.rm = TRUE)} # Function to sum across rows if available, else NA +avg_NA <- function(x) {if (all(is.na(x))) x[NA_integer_] else mean(x, na.rm = TRUE)} # Function to average across rows if available, else NA if (mqc_version == 'old_mqc') { columns <- c( @@ -917,6 +916,13 @@ if (mqc_version == 'old_mqc') { "STAR_mqc.generalstats.star.uniquely_mapped_percent", "featureCounts_mqc.generalstats.featurecounts.percent_assigned" ) + + table_complete <- mqc_stats[,columns] %>% + group_by(Sample) %>% + dplyr::summarize( + across(matches("FastQC_mqc.generalstats.fastqc.total_sequences"), sum_NA), + across(everything(), avg_NA) + ) } else { columns <- c( "Sample", @@ -925,21 +931,28 @@ if (mqc_version == 'old_mqc') { "reads_duplicated_percent" ) + table_complete <- mqc_stats[,columns] %>% + group_by(Sample) %>% + dplyr::summarize( + across(matches("raw_total_sequences"), sum_NA), + across(everything(), avg_NA) + ) + # For the new version, some more tables need to be read if (params$input_type == "salmon" && file.exists(paste0(wd,"/QC/multiqc_data/multiqc_star.txt"))) { star <- read.table(file = paste0(wd,"/QC/multiqc_data/multiqc_star.txt"), header=TRUE, sep="\t", check.names = F)[,c("Sample", "uniquely_mapped_percent")] } else if (params$input_type == "rsem" && file.exists(paste0(wd,"/QC/multiqc_data/multiqc_rsem.txt"))) { - star <- read.table(file = paste0(wd,"/QC/multiqc_data/multiqc_rsem.txt"), header=TRUE, sep="\t", check.names = F)[,c("Sample", "Alignable", "Unique")] - star$uniquely_mapped_percent <- star$Unique/star$Alignable + star <- read.table(file = paste0(wd,"/QC/multiqc_data/multiqc_rsem.txt"), header=TRUE, sep="\t", check.names = F)[,c("Sample", "Total", "Unique")] + star$uniquely_mapped_percent <- star$Unique/star$Total star$Unique <- NULL - star$Alignable <- NULL + star$Total <- NULL } else { star <- NULL } if (file.exists(paste0(wd,"/QC/multiqc_data/multiqc_cutadapt.txt"))) { cutadapt <- read.table(file = paste0(wd,"/QC/multiqc_data/multiqc_cutadapt.txt"), header=TRUE, sep="\t", check.names = F)[,c("Sample", "percent_trimmed")] cutadapt$Sample <- sub("_[0-9]+$", "", cutadapt$Sample) # Remove _1 etc. to produce duplicate rows for summarising - cutadapt <- cutadapt %>% group_by(Sample) %>% summarise_all(sum_NA) + cutadapt <- cutadapt %>% group_by(Sample) %>% summarise_all(avg_NA) } else { cutadapt <- NULL } @@ -952,7 +965,7 @@ if (mqc_version == 'old_mqc') { fastqc_untrimmed <- read.table(file = paste0(wd,"/QC/multiqc_data/multiqc_fastqc.txt"), header=TRUE, sep="\t", check.names = F)[,c("Sample", "%GC")] colnames(fastqc_untrimmed)[colnames(fastqc_untrimmed) == "%GC"] <- "GC_untrimmed" fastqc_untrimmed$Sample <- sub("_[0-9]+$", "", fastqc_untrimmed$Sample) # Remove _1 etc. to produce duplicate rows for summarising - fastqc_untrimmed <- fastqc_untrimmed %>% group_by(Sample) %>% summarise_all(sum_NA) + fastqc_untrimmed <- fastqc_untrimmed %>% group_by(Sample) %>% summarise_all(avg_NA) } else { fastqc_untrimmed <- NULL } @@ -960,14 +973,12 @@ if (mqc_version == 'old_mqc') { fastqc_trimmed <- read.table(file = paste0(wd,"/QC/multiqc_data/multiqc_fastqc_1.txt"), header=TRUE, sep="\t", check.names = F)[,c("Sample", "%GC")] colnames(fastqc_trimmed)[colnames(fastqc_trimmed) == "%GC"] <- "GC_trimmed" fastqc_trimmed$Sample <- sub("_[0-9]+$", "", fastqc_trimmed$Sample) # Remove _1 etc. to produce duplicate rows for summarising - fastqc_trimmed <- fastqc_trimmed %>% group_by(Sample) %>% summarise_all(sum_NA) + fastqc_trimmed <- fastqc_trimmed %>% group_by(Sample) %>% summarise_all(avg_NA) } else { fastqc_trimmed <- NULL } } -table_complete <- mqc_stats[,columns] - if (mqc_version == 'new_mqc') { # Merge the additional tables into the stats From 0789481df0e0d7d4b8ed4ae7b344446e29e0bd23 Mon Sep 17 00:00:00 2001 From: WackerO Date: Mon, 29 Apr 2024 12:54:02 +0200 Subject: [PATCH 35/73] working on removing irrelevant samples from heatmaps_norm --- assets/RNAseq_report.Rmd | 33 ++++++++++++++++++++++++++++++++- 1 file changed, 32 insertions(+), 1 deletion(-) diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index b00bfdec..d990a9d1 100755 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -1885,9 +1885,16 @@ theme_set(theme_classic()) for (file in contrast_files){ - # Reading DE genes list + # Read DE genes list fname <- tools::file_path_sans_ext(basename(file)) + # Find out which conditions are being compared in the current contrast to filter the heatmaps matrix + comparison <- unlist(strsplit(fname, "_", fixed=T)) + vs <- which(comparison == "vs") # Get position of 'vs', e.g. DE_contrast_condition_cellline_H1_vs_H2 --> vs = 6 + cond0 <- comparison[vs-2] # Get contrast, e.g. cellline + cond1 <- comparison[vs-1] # Get first condition, e.g. H1 + cond2 <- comparison[vs+1] # Get second condition, e.g. H2 + dir.create(paste("pathway_analysis", fname, sep="/")) dir.create(paste("pathway_analysis", fname, "KEGG_pathways", sep="/")) dir.create(paste("pathway_analysis", fname, "enrichment_plots", sep="/")) @@ -2050,6 +2057,30 @@ for (file in contrast_files){ mat$gene_name <- NULL mat$Ensembl_ID <- NULL mat <- data.matrix(mat) + capture.output(fname, file="/home-link/iivow01/git/rnadeseq/error/fname", append=F) + capture.output(vs, file="/home-link/iivow01/git/rnadeseq/error/fname", append=T) + capture.output(cond0, file="/home-link/iivow01/git/rnadeseq/error/fname", append=T) + capture.output(cond1, file="/home-link/iivow01/git/rnadeseq/error/fname", append=T) + capture.output(cond2, file="/home-link/iivow01/git/rnadeseq/error/fname", append=T) + write.table(mat, file="/home-link/iivow01/git/rnadeseq/error/mat.tsv", quote=F, sep="\t") + write.table(metadata_save, file="/home-link/iivow01/git/rnadeseq/error/metadata_save.tsv", quote=F, sep="\t") + + # Find metadata column name of the current contrast + cond0_keep <- unlist(grep(paste0("(", cond0, ")$"), colnames(metadata_save), value=T))[1] + capture.output(grep(paste0("(", cond0, ")$"), colnames(metadata_save), value=T), file="/home-link/iivow01/git/rnadeseq/error/grep", append=F) + capture.output(cond0_keep, file="/home-link/iivow01/git/rnadeseq/error/cond0_keep", append=F) + # Subset metadata to those samples that have the current conditions in the current contrast, e.g. genotype is H1 or H2 + metadata_keep <- metadata_save[metadata_save[,cond0_keep] %in% c(cond1,cond2),] + write.table(metadata_keep, file="/home-link/iivow01/git/rnadeseq/error/metadata_keep.tsv", quote=F, sep="\t") +# Subset matrix accordingly + capture.output(rownames(metadata_keep), file="/home-link/iivow01/git/rnadeseq/error/row_met", append=F) + capture.output(colnames(mat), file="/home-link/iivow01/git/rnadeseq/error/cols_mat", append=F) + capture.output(typeof(mat), file="/home-link/iivow01/git/rnadeseq/error/type_mat", append=F) + + mat <- as.data.frame(mat[,rownames(metadata_keep)]) + capture.output(typeof(mat), file="/home-link/iivow01/git/rnadeseq/error/type_mat", append=T) + write.table(mat, file="/home-link/iivow01/git/rnadeseq/error/mat2.tsv", quote=F, sep="\t") + # Skip the heatmaps for CI tests because especially png() works only very unreliably for the heatmaps for some reason if (!isProvided(params$citest) & nrow(mat)>1){ From b01328b9ba97feefd7198314d7283e07d16d9516 Mon Sep 17 00:00:00 2001 From: WackerO Date: Mon, 29 Apr 2024 13:47:27 +0200 Subject: [PATCH 36/73] Undid some changes about sample filtering for heatmaps --- assets/RNAseq_report.Rmd | 31 ------------------------------- 1 file changed, 31 deletions(-) diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index d990a9d1..c52394fe 100755 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -1888,13 +1888,6 @@ for (file in contrast_files){ # Read DE genes list fname <- tools::file_path_sans_ext(basename(file)) - # Find out which conditions are being compared in the current contrast to filter the heatmaps matrix - comparison <- unlist(strsplit(fname, "_", fixed=T)) - vs <- which(comparison == "vs") # Get position of 'vs', e.g. DE_contrast_condition_cellline_H1_vs_H2 --> vs = 6 - cond0 <- comparison[vs-2] # Get contrast, e.g. cellline - cond1 <- comparison[vs-1] # Get first condition, e.g. H1 - cond2 <- comparison[vs+1] # Get second condition, e.g. H2 - dir.create(paste("pathway_analysis", fname, sep="/")) dir.create(paste("pathway_analysis", fname, "KEGG_pathways", sep="/")) dir.create(paste("pathway_analysis", fname, "enrichment_plots", sep="/")) @@ -2057,30 +2050,6 @@ for (file in contrast_files){ mat$gene_name <- NULL mat$Ensembl_ID <- NULL mat <- data.matrix(mat) - capture.output(fname, file="/home-link/iivow01/git/rnadeseq/error/fname", append=F) - capture.output(vs, file="/home-link/iivow01/git/rnadeseq/error/fname", append=T) - capture.output(cond0, file="/home-link/iivow01/git/rnadeseq/error/fname", append=T) - capture.output(cond1, file="/home-link/iivow01/git/rnadeseq/error/fname", append=T) - capture.output(cond2, file="/home-link/iivow01/git/rnadeseq/error/fname", append=T) - write.table(mat, file="/home-link/iivow01/git/rnadeseq/error/mat.tsv", quote=F, sep="\t") - write.table(metadata_save, file="/home-link/iivow01/git/rnadeseq/error/metadata_save.tsv", quote=F, sep="\t") - - # Find metadata column name of the current contrast - cond0_keep <- unlist(grep(paste0("(", cond0, ")$"), colnames(metadata_save), value=T))[1] - capture.output(grep(paste0("(", cond0, ")$"), colnames(metadata_save), value=T), file="/home-link/iivow01/git/rnadeseq/error/grep", append=F) - capture.output(cond0_keep, file="/home-link/iivow01/git/rnadeseq/error/cond0_keep", append=F) - # Subset metadata to those samples that have the current conditions in the current contrast, e.g. genotype is H1 or H2 - metadata_keep <- metadata_save[metadata_save[,cond0_keep] %in% c(cond1,cond2),] - write.table(metadata_keep, file="/home-link/iivow01/git/rnadeseq/error/metadata_keep.tsv", quote=F, sep="\t") -# Subset matrix accordingly - capture.output(rownames(metadata_keep), file="/home-link/iivow01/git/rnadeseq/error/row_met", append=F) - capture.output(colnames(mat), file="/home-link/iivow01/git/rnadeseq/error/cols_mat", append=F) - capture.output(typeof(mat), file="/home-link/iivow01/git/rnadeseq/error/type_mat", append=F) - - mat <- as.data.frame(mat[,rownames(metadata_keep)]) - capture.output(typeof(mat), file="/home-link/iivow01/git/rnadeseq/error/type_mat", append=T) - write.table(mat, file="/home-link/iivow01/git/rnadeseq/error/mat2.tsv", quote=F, sep="\t") - # Skip the heatmaps for CI tests because especially png() works only very unreliably for the heatmaps for some reason if (!isProvided(params$citest) & nrow(mat)>1){ From ce9bf0abfad672896b83c68193e2dee4aff75b7f Mon Sep 17 00:00:00 2001 From: WackerO Date: Mon, 29 Apr 2024 14:33:40 +0200 Subject: [PATCH 37/73] Fixed enrichment plots having their y labels cut off --- assets/RNAseq_report.Rmd | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index c52394fe..ff7f70d3 100755 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -2026,7 +2026,7 @@ for (file in contrast_files){ geom_bar(aes(fill=Padj), stat="identity", width = 0.7) + geom_text(aes(label=paste0(df_subset$DE_genes, "/", df_subset$Pathway_size)), vjust=0.4, hjust=-0.5, size=3) + coord_flip() + - scale_y_continuous(limits = c(0.00, 1.00)) + + scale_y_continuous(limits = c(0.00, 1.00), expand=expansion(mult=c(0,0.2))) + scale_fill_continuous(high = "#132B43", low = "#56B1F7") + ggtitle("Enriched pathways") + xlab("") + ylab("Gene fraction (DE genes / Pathway size)") From dab8978763b35a673033a73c63fdf0a6a417f4ec Mon Sep 17 00:00:00 2001 From: WackerO Date: Mon, 29 Apr 2024 14:41:52 +0200 Subject: [PATCH 38/73] Update changelog --- CHANGELOG.md | 19 ++++++++++++++++--- 1 file changed, 16 insertions(+), 3 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index a9e2e139..9c2ecb7b 100755 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -3,11 +3,26 @@ The format is based on [Keep a Changelog](https://keepachangelog.com/en/1.0.0/) and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0.html). -## 2.2 Avenue of Poplars +## 2.3dev ### Added +- [#229](https://github.com/qbic-pipelines/rnadeseq/pull/229) Added param for clustering (or not) the heatmaps - [#225](https://github.com/qbic-pipelines/rnadeseq/pull/225) Added param for pathway analysis datasources + +### Changed + +- [#225](https://github.com/qbic-pipelines/rnadeseq/pull/225) Shortened names of some output files + +### Fixed + +- [#229](https://github.com/qbic-pipelines/rnadeseq/pull/229) Fixed cutoff enrichment plot labels, fixed wrong plotMA function being called (also fixed this changelog) +- [#225](https://github.com/qbic-pipelines/rnadeseq/pull/225) Fixed too many devices error from tryCatch around normalized heatmaps + +## 2.2 Avenue of Poplars + +### Added + - [#215](https://github.com/qbic-pipelines/rnadeseq/pull/215) Added gene_name to boxplot titles and filenames; increased threshold before overlapping PCA labels are hidden - [#213](https://github.com/qbic-pipelines/rnadeseq/pull/213) Added smrnaseq input support - [#212](https://github.com/qbic-pipelines/rnadeseq/pull/212) Added computational methods if no --software_versions @@ -18,7 +33,6 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### Changed -- [#225](https://github.com/qbic-pipelines/rnadeseq/pull/225) Shortened names of some output files - [#219](https://github.com/qbic-pipelines/rnadeseq/pull/219) Release 2.2 to master - [#220](https://github.com/qbic-pipelines/rnadeseq/pull/220) Commit suggestions from PR review for release 2.2 - [#218](https://github.com/qbic-pipelines/rnadeseq/pull/218) Preparing release 2.2 with version bumps @@ -36,7 +50,6 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### Fixed -- [#225](https://github.com/qbic-pipelines/rnadeseq/pull/225) Fixed too many devices error from tryCatch around normalized heatmaps - [#212](https://github.com/qbic-pipelines/rnadeseq/pull/212) Fixed movability of interactive gostplots - [#208](https://github.com/qbic-pipelines/rnadeseq/pull/208) Fixed relevel bug, the function should now finally work! - [#207](https://github.com/qbic-pipelines/rnadeseq/pull/207) Fixed check of samples in counts vs metadata From cb0c1e4b419b12934030ced922dc7136a485df60 Mon Sep 17 00:00:00 2001 From: WackerO Date: Mon, 29 Apr 2024 15:22:00 +0200 Subject: [PATCH 39/73] updated test profiles --- tests/test.yml | 2 +- tests/test_custom_gmt.yml | 2 +- tests/test_full.yml | 2 +- tests/test_star_rsem.yml | 4 ++-- 4 files changed, 5 insertions(+), 5 deletions(-) diff --git a/tests/test.yml b/tests/test.yml index 585f4a04..40fa1b77 100755 --- a/tests/test.yml +++ b/tests/test.yml @@ -60,6 +60,7 @@ - path: results_test/differential_gene_expression/plots/PCA_plot.pdf - path: results_test/differential_gene_expression/plots/PCA_plot.png - path: results_test/differential_gene_expression/plots/PCA_plot.svg + - path: results_test/differential_gene_expression/plots/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_venn_diagram.pdf - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_venn_diagram.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_venn_diagram.svg @@ -91,5 +92,4 @@ - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/gost_pathway_venn_diagram.svg - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/enrichment_plots/REAC_pathway_enrichment_plot.pdf - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/enrichment_plots/REAC_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf - path: results_test/RNAseq_report.html diff --git a/tests/test_custom_gmt.yml b/tests/test_custom_gmt.yml index ea161a15..baa32547 100755 --- a/tests/test_custom_gmt.yml +++ b/tests/test_custom_gmt.yml @@ -60,6 +60,7 @@ - path: results_test/differential_gene_expression/plots/PCA_plot.pdf - path: results_test/differential_gene_expression/plots/PCA_plot.png - path: results_test/differential_gene_expression/plots/PCA_plot.svg + - path: results_test/differential_gene_expression/plots/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_gostplot.pdf - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_gostplot.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_gostplot.svg @@ -70,5 +71,4 @@ md5sum: eae154d9c7a526cc8c3fcd1a586fa0ed - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/pathway_enrichment_results.tsv md5sum: bd2cbbf85c3e241743e60b9b8263a31c - - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf - path: results_test/RNAseq_report.html diff --git a/tests/test_full.yml b/tests/test_full.yml index beed5c7d..18012af8 100755 --- a/tests/test_full.yml +++ b/tests/test_full.yml @@ -60,6 +60,7 @@ - path: results_test/differential_gene_expression/plots/PCA_plot.pdf - path: results_test/differential_gene_expression/plots/PCA_plot.png - path: results_test/differential_gene_expression/plots/PCA_plot.svg + - path: results_test/differential_gene_expression/plots/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_gostplot.pdf - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/gost_pathway_gostplot.png - path: results_test/pathway_analysis/DE_contrast_condition_genotype_WT_vs_KO/pathway_enrichment_results.tsv @@ -81,5 +82,4 @@ md5sum: 6594fdb8c0eb591fa131982a8be00877 - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/enrichment_plots/REAC_pathway_enrichment_plot.pdf - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/enrichment_plots/REAC_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf - path: results_test/RNAseq_report.html diff --git a/tests/test_star_rsem.yml b/tests/test_star_rsem.yml index d0d4b11c..abd12cb3 100755 --- a/tests/test_star_rsem.yml +++ b/tests/test_star_rsem.yml @@ -75,6 +75,8 @@ - path: results_test/differential_gene_expression/plots/PCA_plot.pdf - path: results_test/differential_gene_expression/plots/PCA_plot.png - path: results_test/differential_gene_expression/plots/PCA_plot.svg + - path: results_test/differential_gene_expression/plots/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf + - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.svg - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.pdf - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.png - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.svg @@ -112,6 +114,4 @@ - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/gost_pathway_venn_diagram.svg - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/pathway_enrichment_results.tsv md5sum: 68f919b58d7fe5cc1b9a38665abccf78 - - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf - - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.svg - path: results_test/pathway_analysis/metadata/gprofiler_full_hsapiens.ENSG.gmt From 41afb0698ba9a2728c4088d9ac9f8917c5a6e4d4 Mon Sep 17 00:00:00 2001 From: WackerO Date: Tue, 30 Apr 2024 08:40:14 +0200 Subject: [PATCH 40/73] fixed multiqc paths in config --- conf/test_star_rsem.config | 2 +- conf/test_star_salmon.config | 2 +- 2 files changed, 2 insertions(+), 2 deletions(-) diff --git a/conf/test_star_rsem.config b/conf/test_star_rsem.config index 840a5191..2139133b 100755 --- a/conf/test_star_rsem.config +++ b/conf/test_star_rsem.config @@ -23,7 +23,7 @@ params { //report_options = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/report_options.yml' project_summary = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/summary.tsv' software_versions = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/software_versions_rsem.yml' - multiqc = 'testdata/QDESQ/new_rsem_multiqc.zip' + multiqc = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/QDESQ/new_rsem_multiqc.zip' input_type = 'rsem' run_pathway_analysis = true datasources = 'GO:CC,KEGG' diff --git a/conf/test_star_salmon.config b/conf/test_star_salmon.config index 708c532c..3fd41063 100755 --- a/conf/test_star_salmon.config +++ b/conf/test_star_salmon.config @@ -23,7 +23,7 @@ params { //report_options = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/report_options.yml' project_summary = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/summary.tsv' software_versions = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/software_versions_salmon.yml' - multiqc = 'testdata/QDESQ/new_salmon_multiqc.zip' + multiqc = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/QDESQ/new_salmon_multiqc.zip' input_type = 'salmon' run_pathway_analysis = true datasources = 'KEGG,REAC' From fb9a4bd854a710f70ab32365848e135d5ed98e36 Mon Sep 17 00:00:00 2001 From: WackerO Date: Tue, 30 Apr 2024 08:44:35 +0200 Subject: [PATCH 41/73] fixed heatmap test path --- tests/test_star_rsem.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/tests/test_star_rsem.yml b/tests/test_star_rsem.yml index abd12cb3..3aba2020 100755 --- a/tests/test_star_rsem.yml +++ b/tests/test_star_rsem.yml @@ -76,7 +76,7 @@ - path: results_test/differential_gene_expression/plots/PCA_plot.png - path: results_test/differential_gene_expression/plots/PCA_plot.svg - path: results_test/differential_gene_expression/plots/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf - - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.svg + - path: results_test/differential_gene_expression/heatmap_gene_list/Heatmap_normalized_counts_gene_list.svg - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.pdf - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.png - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.svg From f1edb412330b0212e3b6c1c6c5cda6a742177a46 Mon Sep 17 00:00:00 2001 From: WackerO Date: Tue, 30 Apr 2024 13:14:23 +0200 Subject: [PATCH 42/73] fixed test profile --- tests/test_star_rsem.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/tests/test_star_rsem.yml b/tests/test_star_rsem.yml index 3aba2020..d1f39921 100755 --- a/tests/test_star_rsem.yml +++ b/tests/test_star_rsem.yml @@ -76,7 +76,7 @@ - path: results_test/differential_gene_expression/plots/PCA_plot.png - path: results_test/differential_gene_expression/plots/PCA_plot.svg - path: results_test/differential_gene_expression/plots/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf - - path: results_test/differential_gene_expression/heatmap_gene_list/Heatmap_normalized_counts_gene_list.svg + - path: results_test/differential_gene_expression/plots/heatmap_gene_list/Heatmap_normalized_counts_gene_list.svg - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.pdf - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.png - path: results_test/pathway_analysis/DE_contrast_condition_cellline_H1_vs_GM12878/enrichment_plots/GO.CC_pathway_enrichment_plot.svg From e3a8f66c4d9f00e2c05fc1a56c773449eabc5151 Mon Sep 17 00:00:00 2001 From: WackerO Date: Tue, 30 Apr 2024 15:15:54 +0200 Subject: [PATCH 43/73] fixed md5sum --- tests/test_star_rsem.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/tests/test_star_rsem.yml b/tests/test_star_rsem.yml index d1f39921..59120301 100755 --- a/tests/test_star_rsem.yml +++ b/tests/test_star_rsem.yml @@ -113,5 +113,5 @@ - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/gost_pathway_venn_diagram.png - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/gost_pathway_venn_diagram.svg - path: results_test/pathway_analysis/DE_contrast_condition_cellline_MCF7_vs_GM12878/pathway_enrichment_results.tsv - md5sum: 68f919b58d7fe5cc1b9a38665abccf78 + md5sum: b7f67bc86475b218965b173b753199b1 - path: results_test/pathway_analysis/metadata/gprofiler_full_hsapiens.ENSG.gmt From a5725c9daa453172730ec9423119d1d71a1ec040 Mon Sep 17 00:00:00 2001 From: WackerO Date: Thu, 2 May 2024 14:05:37 +0200 Subject: [PATCH 44/73] Fixed some things from https://github.com/qbic-pipelines/rnadeseq/pull/226 --- assets/RNAseq_report.Rmd | 40 +++++++++++++++++++++------------------- 1 file changed, 21 insertions(+), 19 deletions(-) diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index 500fc7f5..b16c0ea7 100755 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -869,6 +869,17 @@ for (i in resultsNames(cds)[-1]) { ```{r summary_stats, echo=FALSE, message=FALSE, warning=FALSE, results='asis', eval=dir.exists(paste0(wd,"/QC/multiqc_data/")), results = 'asis'} +# Check first if a new or old multiqc file was provided +if (file.exists(paste0(wd,"/QC/multiqc_data/multiqc_general_stats.txt"))) { + mqc_stats <- read.table(file = paste0(wd,"/QC/multiqc_data/multiqc_general_stats.txt"), header=TRUE, sep="\t") + mqc_version <- 'old_mqc' +} else if (file.exists(paste0(wd,"/QC/multiqc_data/multiqc_samtools_stats.txt"))) { + mqc_stats <- read.table(file = paste0(wd,"/QC/multiqc_data/multiqc_samtools_stats.txt"), header=TRUE, sep="\t") + mqc_version <- 'new_mqc' +} else { + stop("Could not find a suitable multiqc table; please provide a correct multiqc.zip file or omit the parameter altogether") +} + cat(paste0("*** # Read mapping and assignment @@ -880,20 +891,12 @@ The read sequencing quality was analyzed using `FASTQC`. More detailed informati The read mapping and gene assignment statistics, together with other quality control parameters are summarized here. -The table below, extracted from the `MultiQC` report, shows a summary of the bioinformatics analysis quality control. -Note: Duplicate intercept (%) is the percentage of duplicate reads for the intercept of a linear model of duplicate reads vs read counts. High numbers of duplicates at low read counts can indicate low library complexity with technical duplication." +The table below, extracted from the `MultiQC` report, shows a summary of the bioinformatics analysis quality control.", +ifelse(mqc_version == 'old_mqc', +"Note: Duplicate intercept (%) is the percentage of duplicate reads for the intercept of a linear model of duplicate reads vs read counts. High numbers of duplicates at low read counts can indicate low library complexity with technical duplication.", +"") )) -if (file.exists(paste0(wd,"/QC/multiqc_data/multiqc_general_stats.txt"))) { - mqc_stats <- read.table(file = paste0(wd,"/QC/multiqc_data/multiqc_general_stats.txt"), header=TRUE, sep="\t") - mqc_version <- 'old_mqc' -} else if (file.exists(paste0(wd,"/QC/multiqc_data/multiqc_samtools_stats.txt"))) { - mqc_stats <- read.table(file = paste0(wd,"/QC/multiqc_data/multiqc_samtools_stats.txt"), header=TRUE, sep="\t") - mqc_version <- 'new_mqc' -} else { - stop("Could not find a suitable multiqc table; please provide a correct multiqc.zip file or omit the parameter altogether") -} - is.num <- sapply(mqc_stats, is.numeric) mqc_stats[is.num] <- lapply(mqc_stats[is.num], round, 2) @@ -1007,7 +1010,7 @@ if (mqc_version == 'old_mqc') { table_complete <- table_complete %>% mutate( Sample = Sample, - ReadNumber = color_bar("lightblue")(round((ReadNumber/1000000),2)), + ReadNumber = round((ReadNumber/1000000),2), DuplicateReadsIntercept = ifelse(DuplicateReadsIntercept > 1, cell_spec(DuplicateReadsIntercept, color="orange", bold=T), cell_spec(DuplicateReadsIntercept, color="black")), @@ -1022,7 +1025,6 @@ if (mqc_version == 'old_mqc') { ) } else { colnames <- c("Sample", "Number of reads (M)", "Duplicates (%)", "Untrimmed GC (%)", "Trimmed GC (%)", "Trimmed reads (%)", "Mapped reads (%)", "Assigned reads (%)") - #colnames(table_complete) <- c("Sample", "ReadNumber", "DuplicateReadsIntercept") table_complete <- table_complete %>% transmute( @@ -1037,9 +1039,9 @@ if (mqc_version == 'old_mqc') { MappedReads = ifelse(uniquely_mapped_percent > 80, color_bar("lightblue")(uniquely_mapped_percent), color_bar("orange")(uniquely_mapped_percent)), - AssignedReads = ifelse(total_assigned_tags/total_tags > 60, - color_bar("lightblue")(total_assigned_tags/total_tags), - color_bar("orange")(total_assigned_tags/total_tags)) + AssignedReads = ifelse((total_assigned_tags/total_tags*100) > 60, + color_bar("lightblue")((total_assigned_tags/total_tags)*100), + color_bar("orange")((total_assigned_tags/total_tags)*100)) ) } @@ -1103,9 +1105,9 @@ The statistics of read assignment to genes are shown below. Most reads should be

", " -## Read alignment statistics {.tabset} +## Read assignment statistics {.tabset} -The statistics of read alignment to genes are shown below for pipeline runs on data produced by `rnaseq <= v1.4.2`. If a later version was used, these plots are not generated by rnaseq anymore. However, you can find `star/featurecounts/.featureCounts.txt.summary` or `star_salmon/featurecounts/.featureCounts.txt.summary` files in the rnaseq output folder which contain the necessary information for creating such plots. +The statistics of read assignment to genes are shown below for pipeline runs on data produced by `rnaseq <= v1.4.2`. If a later version was used, these plots are not generated by rnaseq anymore. However, you can find `star/featurecounts/.featureCounts.txt.summary` or `star_salmon/featurecounts/.featureCounts.txt.summary` files in the rnaseq output folder which contain the necessary information for creating such plots. "))) ``` From 18b6b8a774e0f7df702d0447dd60cecc68af03d5 Mon Sep 17 00:00:00 2001 From: WackerO Date: Thu, 2 May 2024 14:06:59 +0200 Subject: [PATCH 45/73] fixed venn paths --- tests/test_full.yml | 6 +++--- 1 file changed, 3 insertions(+), 3 deletions(-) diff --git a/tests/test_full.yml b/tests/test_full.yml index d455e016..5d1beff5 100755 --- a/tests/test_full.yml +++ b/tests/test_full.yml @@ -83,7 +83,7 @@ - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/enrichment_plots/REAC_pathway_enrichment_plot.pdf - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/enrichment_plots/REAC_pathway_enrichment_plot.png - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.pdf - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.png - - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control_gost_pathway_venn_diagram.svg + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/gost_pathway_venn_diagram.pdf + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/gost_pathway_venn_diagram.png + - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/gost_pathway_venn_diagram.svg - path: results_test/RNAseq_report.html From f5b1da2181eda95678326e67d8fc5a8df7ea87e1 Mon Sep 17 00:00:00 2001 From: WackerO Date: Fri, 3 May 2024 08:15:58 +0200 Subject: [PATCH 46/73] removed redundant heatmap from test_full.yml --- tests/test_full.yml | 1 - 1 file changed, 1 deletion(-) diff --git a/tests/test_full.yml b/tests/test_full.yml index 5d1beff5..c44f2338 100755 --- a/tests/test_full.yml +++ b/tests/test_full.yml @@ -82,7 +82,6 @@ md5sum: 6594fdb8c0eb591fa131982a8be00877 - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/enrichment_plots/REAC_pathway_enrichment_plot.pdf - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/enrichment_plots/REAC_pathway_enrichment_plot.png - - path: results_test/pathway_analysis/heatmap_gene_list/Heatmap_normalized_counts_gene_list.pdf - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/gost_pathway_venn_diagram.pdf - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/gost_pathway_venn_diagram.png - path: results_test/pathway_analysis/DE_contrast_condition_treatment_Treated_vs_Control/gost_pathway_venn_diagram.svg From a1e0a932fa92b7f966bfc6922653e4bc970d3e59 Mon Sep 17 00:00:00 2001 From: WackerO Date: Fri, 3 May 2024 12:56:47 +0200 Subject: [PATCH 47/73] Updated Changelog --- CHANGELOG.md | 2 ++ 1 file changed, 2 insertions(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index 65063ff1..3491afdd 100755 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -8,6 +8,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### Added - [#229](https://github.com/qbic-pipelines/rnadeseq/pull/229) Added param for clustering (or not) the heatmaps +- [#226](https://github.com/qbic-pipelines/rnadeseq/pull/226) Added logic to read newer multiqc files from rnaseq 3.12 - [#225](https://github.com/qbic-pipelines/rnadeseq/pull/225) Added param for pathway analysis datasources - [#221](https://github.com/qbic-pipelines/rnadeseq/pull/221) Added padj to volcano hovertext @@ -17,6 +18,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### Fixed +- [#228](https://github.com/qbic-pipelines/rnadeseq/pull/228) Fixed text in report - [#229](https://github.com/qbic-pipelines/rnadeseq/pull/229) Fixed cutoff enrichment plot labels, fixed wrong plotMA function being called (also fixed this changelog) - [#225](https://github.com/qbic-pipelines/rnadeseq/pull/225) Fixed too many devices error from tryCatch around normalized heatmaps - [#221](https://github.com/qbic-pipelines/rnadeseq/pull/221) Fixed non-conformable arrays bug, fix wrong volcano colors when no DE genes From 48d3121cfd4fcadef2aee1b24459f0ef8d98daf0 Mon Sep 17 00:00:00 2001 From: WackerO Date: Mon, 6 May 2024 14:29:52 +0200 Subject: [PATCH 48/73] linebreak change to restart the github ci tests --- assets/RNAseq_report.Rmd | 2 -- 1 file changed, 2 deletions(-) diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index 88c90a8e..3717035a 100755 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -416,8 +416,6 @@ if (params$input_type == "smrnaseq") { count.table$Ensembl_ID <- rownames(count.table) count.table <- cbind(Ensembl_ID=rownames(count.table), gene_name=rownames(count.table), count.table) write.table(count.table, paste("differential_gene_expression/gene_counts_tables/raw_gene_counts.tsv",sep=""), append = FALSE, quote = FALSE, sep = "\t",eol = "\n", na = "NA", dec = ".", row.names = F, qmethod = c("escape", "double")) - - } # Load count table for FeatureCounts From 9fdf6da395d79e5a4cd055e967c75e9d0b95e71c Mon Sep 17 00:00:00 2001 From: WackerO Date: Tue, 7 May 2024 15:28:23 +0200 Subject: [PATCH 49/73] Changed dev to 2.3 in preparattion of release --- .github/workflows/ci.yml | 8 ++++---- Dockerfile | 6 +++--- environment.yml | 2 +- modules/local/report.nf | 2 +- nextflow.config | 2 +- 5 files changed, 10 insertions(+), 10 deletions(-) diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index d1e1401f..ef1b2141 100755 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -40,14 +40,14 @@ jobs: environment.yml - name: Build new docker image if: env.MATCHED_FILES - run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:dev + run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:2.3 # Change the version above and the third version below before/after release - name: Pull docker image if: ${{ !env.MATCHED_FILES }} run: | docker pull ghcr.io/qbic-pipelines/rnadeseq:dev - docker tag ghcr.io/qbic-pipelines/rnadeseq:dev ghcr.io/qbic-pipelines/rnadeseq:dev + docker tag ghcr.io/qbic-pipelines/rnadeseq:dev ghcr.io/qbic-pipelines/rnadeseq:2.3 - name: Install Nextflow uses: nf-core/setup-nextflow@v1 @@ -94,14 +94,14 @@ jobs: environment.yml - name: Build new docker image if: env.MATCHED_FILES - run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:dev + run: docker build --no-cache . -t ghcr.io/qbic-pipelines/rnadeseq:2.3 # Change the version above and the third version below before/after release - name: Pull docker image if: ${{ !env.MATCHED_FILES }} run: | docker pull ghcr.io/qbic-pipelines/rnadeseq:dev - docker tag ghcr.io/qbic-pipelines/rnadeseq:dev ghcr.io/qbic-pipelines/rnadeseq:dev + docker tag ghcr.io/qbic-pipelines/rnadeseq:dev ghcr.io/qbic-pipelines/rnadeseq:2.3 - name: Install Nextflow uses: nf-core/setup-nextflow@v1 diff --git a/Dockerfile b/Dockerfile index 4f8ce7fe..617a13e0 100755 --- a/Dockerfile +++ b/Dockerfile @@ -5,14 +5,14 @@ LABEL org.opencontainers.image.authors="Gisela Gabernet, Alexander Peltzer, Oska LABEL org.opencontainers.image.licenses=MIT COPY environment.yml / #RUN conda install -c conda-forge mamba -RUN mamba env create --file /environment.yml -p /opt/conda/envs/qbic-pipelines-rnadeseq-dev && \ +RUN mamba env create --file /environment.yml -p /opt/conda/envs/qbic-pipelines-rnadeseq-2.3 && \ mamba clean --all --yes RUN apt-get update -qq && \ apt-get install -y zip procps ghostscript # Add conda installation dir to PATH -ENV PATH /opt/conda/envs/qbic-pipelines-rnadeseq-dev/bin:$PATH +ENV PATH /opt/conda/envs/qbic-pipelines-rnadeseq-2.3/bin:$PATH # Dump the details of the installed packates to a file for posterity -RUN mamba env export --name qbic-pipelines-rnadeseq-dev > qbic-pipelines-rnadeseq-dev.yml +RUN mamba env export --name qbic-pipelines-rnadeseq-2.3 > qbic-pipelines-rnadeseq-2.3.yml # Instruct R processes to use these empty files instead of clashing with a local config RUN touch .Rprofile RUN touch .Renviron diff --git a/environment.yml b/environment.yml index cede896c..c759c019 100755 --- a/environment.yml +++ b/environment.yml @@ -1,7 +1,7 @@ # You can use this file to create a conda environment for this pipeline: # conda env create -f environment.yml # use this to find packages: https://anaconda.org/ -name: qbic-pipelines-rnadeseq-dev +name: qbic-pipelines-rnadeseq-2.3 channels: - bioconda - conda-forge diff --git a/modules/local/report.nf b/modules/local/report.nf index 28d8f709..413e241c 100755 --- a/modules/local/report.nf +++ b/modules/local/report.nf @@ -1,6 +1,6 @@ process REPORT { - container 'ghcr.io/qbic-pipelines/rnadeseq:dev' + container 'ghcr.io/qbic-pipelines/rnadeseq:2.3' input: path gene_counts diff --git a/nextflow.config b/nextflow.config index 2026ab6a..4e0c293b 100755 --- a/nextflow.config +++ b/nextflow.config @@ -260,7 +260,7 @@ manifest { description = """Differential gene expression analysis and pathway analysis of RNAseq data""" mainScript = 'main.nf' nextflowVersion = '!>=23.04.0' - version = 'dev' + version = '2.3' doi = '' } From 82f0faebf3bc8e35ea699abbc949cbc25152a7e3 Mon Sep 17 00:00:00 2001 From: WackerO Date: Mon, 13 May 2024 08:54:06 +0200 Subject: [PATCH 50/73] Undid permission changes from previous PR --- .devcontainer/devcontainer.json | 0 .editorconfig | 0 .gitattributes | 0 .github/.dockstore.yml | 0 .github/CONTRIBUTING.md | 0 .github/ISSUE_TEMPLATE/bug_report.yml | 0 .github/ISSUE_TEMPLATE/feature_request.yml | 0 .github/PULL_REQUEST_TEMPLATE.md | 0 .github/workflows/awsfulltest.yml | 0 .github/workflows/awstest.yml | 0 .github/workflows/branch.yml | 0 .github/workflows/ci.yml | 0 .github/workflows/clean-up.yml | 0 .github/workflows/fix-linting.yml | 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mode 100644 diff --git a/tests/test.yml b/tests/test.yml old mode 100755 new mode 100644 diff --git a/tests/test_batcheffect.yml b/tests/test_batcheffect.yml old mode 100755 new mode 100644 diff --git a/tests/test_contrast_list.yml b/tests/test_contrast_list.yml old mode 100755 new mode 100644 diff --git a/tests/test_contrast_matrix.yml b/tests/test_contrast_matrix.yml old mode 100755 new mode 100644 diff --git a/tests/test_custom_gmt.yml b/tests/test_custom_gmt.yml old mode 100755 new mode 100644 diff --git a/tests/test_full.yml b/tests/test_full.yml old mode 100755 new mode 100644 diff --git a/tests/test_no_multiqc.yml b/tests/test_no_multiqc.yml old mode 100755 new mode 100644 diff --git a/tests/test_relevel.yml b/tests/test_relevel.yml old mode 100755 new mode 100644 diff --git a/tests/test_smrnaseq.yml b/tests/test_smrnaseq.yml old mode 100755 new mode 100644 diff --git a/tests/test_star_rsem.yml b/tests/test_star_rsem.yml old mode 100755 new mode 100644 diff --git a/tests/test_star_salmon.yml b/tests/test_star_salmon.yml old mode 100755 new mode 100644 diff --git a/tests/test_use_vst.yml b/tests/test_use_vst.yml old mode 100755 new mode 100644 diff --git a/tower.yml b/tower.yml old mode 100755 new mode 100644 diff --git a/workflows/rnadeseq.nf b/workflows/rnadeseq.nf old mode 100755 new mode 100644 From bc2e18a8e06b42a1527cdd9bd46dbbee34e60af1 Mon Sep 17 00:00:00 2001 From: WackerO Date: Tue, 14 May 2024 11:09:00 +0200 Subject: [PATCH 51/73] updated changelog --- CHANGELOG.md | 1 + 1 file changed, 1 insertion(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index 3491afdd..87d54291 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -18,6 +18,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### Fixed +- [#234](https://github.com/qbic-pipelines/rnadeseq/pull/234) Fixed unnecessary file permission changes from #225 - [#228](https://github.com/qbic-pipelines/rnadeseq/pull/228) Fixed text in report - [#229](https://github.com/qbic-pipelines/rnadeseq/pull/229) Fixed cutoff enrichment plot labels, fixed wrong plotMA function being called (also fixed this changelog) - [#225](https://github.com/qbic-pipelines/rnadeseq/pull/225) Fixed too many devices error from tryCatch around normalized heatmaps From 004c78a484de33b91904da170660c5fbc8ac00fe Mon Sep 17 00:00:00 2001 From: WackerO Date: Wed, 15 May 2024 11:31:16 +0200 Subject: [PATCH 52/73] Initial template commit --- .devcontainer/devcontainer.json | 20 + .editorconfig | 33 ++ .gitattributes | 4 + .github/.dockstore.yml | 6 + .github/CONTRIBUTING.md | 125 +++++ .github/ISSUE_TEMPLATE/bug_report.yml | 50 ++ .github/ISSUE_TEMPLATE/config.yml | 7 + .github/ISSUE_TEMPLATE/feature_request.yml | 11 + .github/PULL_REQUEST_TEMPLATE.md | 26 + .github/workflows/awsfulltest.yml | 39 ++ .github/workflows/awstest.yml | 33 ++ .github/workflows/branch.yml | 44 ++ .github/workflows/ci.yml | 46 ++ .github/workflows/clean-up.yml | 24 + .github/workflows/download_pipeline.yml | 86 ++++ .github/workflows/fix-linting.yml | 89 ++++ .github/workflows/linting.yml | 68 +++ .github/workflows/linting_comment.yml | 28 ++ .github/workflows/release-announcements.yml | 75 +++ .gitignore | 8 + .gitpod.yml | 20 + .nf-core.yml | 2 + .pre-commit-config.yaml | 13 + .prettierignore | 12 + .prettierrc.yml | 1 + CHANGELOG.md | 16 + CITATIONS.md | 41 ++ CODE_OF_CONDUCT.md | 182 +++++++ LICENSE | 21 + README.md | 110 +++++ assets/adaptivecard.json | 67 +++ assets/email_template.html | 53 +++ assets/email_template.txt | 39 ++ assets/methods_description_template.yml | 29 ++ assets/multiqc_config.yml | 15 + assets/nf-core-rnadeseq_logo_light.png | Bin 0 -> 78987 bytes assets/samplesheet.csv | 3 + assets/schema_input.json | 33 ++ assets/sendmail_template.txt | 53 +++ assets/slackreport.json | 34 ++ conf/base.config | 62 +++ conf/igenomes.config | 440 +++++++++++++++++ conf/modules.config | 34 ++ conf/test.config | 29 ++ conf/test_full.config | 24 + docs/README.md | 10 + docs/images/mqc_fastqc_adapter.png | Bin 0 -> 23458 bytes docs/images/mqc_fastqc_counts.png | Bin 0 -> 33918 bytes docs/images/mqc_fastqc_quality.png | Bin 0 -> 55769 bytes docs/images/nf-core-rnadeseq_logo_dark.png | Bin 0 -> 29530 bytes docs/images/nf-core-rnadeseq_logo_light.png | Bin 0 -> 25217 bytes docs/output.md | 71 +++ docs/usage.md | 226 +++++++++ main.nf | 112 +++++ modules.json | 41 ++ modules/nf-core/fastqc/environment.yml | 7 + modules/nf-core/fastqc/main.nf | 61 +++ modules/nf-core/fastqc/meta.yml | 57 +++ modules/nf-core/fastqc/tests/main.nf.test | 212 +++++++++ .../nf-core/fastqc/tests/main.nf.test.snap | 88 ++++ modules/nf-core/fastqc/tests/tags.yml | 2 + modules/nf-core/multiqc/environment.yml | 7 + modules/nf-core/multiqc/main.nf | 55 +++ modules/nf-core/multiqc/meta.yml | 58 +++ modules/nf-core/multiqc/tests/main.nf.test | 84 ++++ .../nf-core/multiqc/tests/main.nf.test.snap | 41 ++ modules/nf-core/multiqc/tests/tags.yml | 2 + nextflow.config | 280 +++++++++++ nextflow_schema.json | 296 ++++++++++++ .../utils_nfcore_rnadeseq_pipeline/main.nf | 263 +++++++++++ .../nf-core/utils_nextflow_pipeline/main.nf | 126 +++++ .../nf-core/utils_nextflow_pipeline/meta.yml | 38 ++ .../tests/main.function.nf.test | 54 +++ .../tests/main.function.nf.test.snap | 20 + .../tests/main.workflow.nf.test | 111 +++++ .../tests/nextflow.config | 9 + .../utils_nextflow_pipeline/tests/tags.yml | 2 + .../nf-core/utils_nfcore_pipeline/main.nf | 446 ++++++++++++++++++ .../nf-core/utils_nfcore_pipeline/meta.yml | 24 + .../tests/main.function.nf.test | 134 ++++++ .../tests/main.function.nf.test.snap | 166 +++++++ .../tests/main.workflow.nf.test | 29 ++ .../tests/main.workflow.nf.test.snap | 19 + .../tests/nextflow.config | 9 + .../utils_nfcore_pipeline/tests/tags.yml | 2 + .../nf-core/utils_nfvalidation_plugin/main.nf | 62 +++ .../utils_nfvalidation_plugin/meta.yml | 44 ++ .../tests/main.nf.test | 200 ++++++++ .../tests/nextflow_schema.json | 96 ++++ .../utils_nfvalidation_plugin/tests/tags.yml | 2 + tower.yml | 5 + workflows/rnadeseq.nf | 98 ++++ 92 files changed, 5824 insertions(+) create mode 100644 .devcontainer/devcontainer.json create mode 100644 .editorconfig create mode 100644 .gitattributes create mode 100644 .github/.dockstore.yml create mode 100644 .github/CONTRIBUTING.md create mode 100644 .github/ISSUE_TEMPLATE/bug_report.yml create mode 100644 .github/ISSUE_TEMPLATE/config.yml create mode 100644 .github/ISSUE_TEMPLATE/feature_request.yml create mode 100644 .github/PULL_REQUEST_TEMPLATE.md create mode 100644 .github/workflows/awsfulltest.yml create mode 100644 .github/workflows/awstest.yml create mode 100644 .github/workflows/branch.yml create mode 100644 .github/workflows/ci.yml create mode 100644 .github/workflows/clean-up.yml create mode 100644 .github/workflows/download_pipeline.yml create mode 100644 .github/workflows/fix-linting.yml create mode 100644 .github/workflows/linting.yml create mode 100644 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100644 modules/nf-core/multiqc/main.nf create mode 100644 modules/nf-core/multiqc/meta.yml create mode 100644 modules/nf-core/multiqc/tests/main.nf.test create mode 100644 modules/nf-core/multiqc/tests/main.nf.test.snap create mode 100644 modules/nf-core/multiqc/tests/tags.yml create mode 100644 nextflow.config create mode 100644 nextflow_schema.json create mode 100644 subworkflows/local/utils_nfcore_rnadeseq_pipeline/main.nf create mode 100644 subworkflows/nf-core/utils_nextflow_pipeline/main.nf create mode 100644 subworkflows/nf-core/utils_nextflow_pipeline/meta.yml create mode 100644 subworkflows/nf-core/utils_nextflow_pipeline/tests/main.function.nf.test create mode 100644 subworkflows/nf-core/utils_nextflow_pipeline/tests/main.function.nf.test.snap create mode 100644 subworkflows/nf-core/utils_nextflow_pipeline/tests/main.workflow.nf.test create mode 100644 subworkflows/nf-core/utils_nextflow_pipeline/tests/nextflow.config create mode 100644 subworkflows/nf-core/utils_nextflow_pipeline/tests/tags.yml create mode 100644 subworkflows/nf-core/utils_nfcore_pipeline/main.nf create mode 100644 subworkflows/nf-core/utils_nfcore_pipeline/meta.yml create mode 100644 subworkflows/nf-core/utils_nfcore_pipeline/tests/main.function.nf.test create mode 100644 subworkflows/nf-core/utils_nfcore_pipeline/tests/main.function.nf.test.snap create mode 100644 subworkflows/nf-core/utils_nfcore_pipeline/tests/main.workflow.nf.test create mode 100644 subworkflows/nf-core/utils_nfcore_pipeline/tests/main.workflow.nf.test.snap create mode 100644 subworkflows/nf-core/utils_nfcore_pipeline/tests/nextflow.config create mode 100644 subworkflows/nf-core/utils_nfcore_pipeline/tests/tags.yml create mode 100644 subworkflows/nf-core/utils_nfvalidation_plugin/main.nf create mode 100644 subworkflows/nf-core/utils_nfvalidation_plugin/meta.yml create mode 100644 subworkflows/nf-core/utils_nfvalidation_plugin/tests/main.nf.test create mode 100644 subworkflows/nf-core/utils_nfvalidation_plugin/tests/nextflow_schema.json create mode 100644 subworkflows/nf-core/utils_nfvalidation_plugin/tests/tags.yml create mode 100644 tower.yml create mode 100644 workflows/rnadeseq.nf diff --git a/.devcontainer/devcontainer.json b/.devcontainer/devcontainer.json new file mode 100644 index 00000000..b290e090 --- /dev/null +++ b/.devcontainer/devcontainer.json @@ -0,0 +1,20 @@ +{ + "name": "nfcore", + "image": "nfcore/gitpod:latest", + "remoteUser": "gitpod", + "runArgs": ["--privileged"], + + // Configure tool-specific properties. + "customizations": { + // Configure properties specific to VS Code. + "vscode": { + // Set *default* container specific settings.json values on container create. + "settings": { + "python.defaultInterpreterPath": "/opt/conda/bin/python" + }, + + // Add the IDs of extensions you want installed when the container is created. + "extensions": ["ms-python.python", "ms-python.vscode-pylance", "nf-core.nf-core-extensionpack"] + } + } +} diff --git a/.editorconfig b/.editorconfig new file mode 100644 index 00000000..72dda289 --- /dev/null +++ b/.editorconfig @@ -0,0 +1,33 @@ +root = true + +[*] +charset = utf-8 +end_of_line = lf +insert_final_newline = true +trim_trailing_whitespace = true +indent_size = 4 +indent_style = space + +[*.{md,yml,yaml,html,css,scss,js}] +indent_size = 2 + +# These files are edited and tested upstream in nf-core/modules +[/modules/nf-core/**] +charset = unset +end_of_line = unset +insert_final_newline = unset +trim_trailing_whitespace = unset +indent_style = unset +[/subworkflows/nf-core/**] +charset = unset +end_of_line = unset +insert_final_newline = unset +trim_trailing_whitespace = unset +indent_style = unset + +[/assets/email*] +indent_size = unset + +# ignore python and markdown +[*.{py,md}] +indent_style = unset diff --git a/.gitattributes b/.gitattributes new file mode 100644 index 00000000..7a2dabc2 --- /dev/null +++ b/.gitattributes @@ -0,0 +1,4 @@ +*.config linguist-language=nextflow +*.nf.test linguist-language=nextflow +modules/nf-core/** linguist-generated +subworkflows/nf-core/** linguist-generated diff --git a/.github/.dockstore.yml b/.github/.dockstore.yml new file mode 100644 index 00000000..191fabd2 --- /dev/null +++ b/.github/.dockstore.yml @@ -0,0 +1,6 @@ +# Dockstore config version, not pipeline version +version: 1.2 +workflows: + - subclass: nfl + primaryDescriptorPath: /nextflow.config + publish: True diff --git a/.github/CONTRIBUTING.md b/.github/CONTRIBUTING.md new file mode 100644 index 00000000..811b1f55 --- /dev/null +++ b/.github/CONTRIBUTING.md @@ -0,0 +1,125 @@ +# nf-core/rnadeseq: Contributing Guidelines + +Hi there! +Many thanks for taking an interest in improving nf-core/rnadeseq. + +We try to manage the required tasks for nf-core/rnadeseq using GitHub issues, you probably came to this page when creating one. +Please use the pre-filled template to save time. + +However, don't be put off by this template - other more general issues and suggestions are welcome! +Contributions to the code are even more welcome ;) + +> [!NOTE] +> If you need help using or modifying nf-core/rnadeseq then the best place to ask is on the nf-core Slack [#rnadeseq](https://nfcore.slack.com/channels/rnadeseq) channel ([join our Slack here](https://nf-co.re/join/slack)). + +## Contribution workflow + +If you'd like to write some code for nf-core/rnadeseq, the standard workflow is as follows: + +1. Check that there isn't already an issue about your idea in the [nf-core/rnadeseq issues](https://github.com/nf-core/rnadeseq/issues) to avoid duplicating work. If there isn't one already, please create one so that others know you're working on this +2. [Fork](https://help.github.com/en/github/getting-started-with-github/fork-a-repo) the [nf-core/rnadeseq repository](https://github.com/nf-core/rnadeseq) to your GitHub account +3. Make the necessary changes / additions within your forked repository following [Pipeline conventions](#pipeline-contribution-conventions) +4. Use `nf-core schema build` and add any new parameters to the pipeline JSON schema (requires [nf-core tools](https://github.com/nf-core/tools) >= 1.10). +5. Submit a Pull Request against the `dev` branch and wait for the code to be reviewed and merged + +If you're not used to this workflow with git, you can start with some [docs from GitHub](https://help.github.com/en/github/collaborating-with-issues-and-pull-requests) or even their [excellent `git` resources](https://try.github.io/). + +## Tests + +You have the option to test your changes locally by running the pipeline. For receiving warnings about process selectors and other `debug` information, it is recommended to use the debug profile. Execute all the tests with the following command: + +```bash +nf-test test --profile debug,test,docker --verbose +``` + +When you create a pull request with changes, [GitHub Actions](https://github.com/features/actions) will run automatic tests. +Typically, pull-requests are only fully reviewed when these tests are passing, though of course we can help out before then. + +There are typically two types of tests that run: + +### Lint tests + +`nf-core` has a [set of guidelines](https://nf-co.re/developers/guidelines) which all pipelines must adhere to. +To enforce these and ensure that all pipelines stay in sync, we have developed a helper tool which runs checks on the pipeline code. This is in the [nf-core/tools repository](https://github.com/nf-core/tools) and once installed can be run locally with the `nf-core lint ` command. + +If any failures or warnings are encountered, please follow the listed URL for more documentation. + +### Pipeline tests + +Each `nf-core` pipeline should be set up with a minimal set of test-data. +`GitHub Actions` then runs the pipeline on this data to ensure that it exits successfully. +If there are any failures then the automated tests fail. +These tests are run both with the latest available version of `Nextflow` and also the minimum required version that is stated in the pipeline code. + +## Patch + +:warning: Only in the unlikely and regretful event of a release happening with a bug. + +- On your own fork, make a new branch `patch` based on `upstream/master`. +- Fix the bug, and bump version (X.Y.Z+1). +- A PR should be made on `master` from patch to directly this particular bug. + +## Getting help + +For further information/help, please consult the [nf-core/rnadeseq documentation](https://nf-co.re/rnadeseq/usage) and don't hesitate to get in touch on the nf-core Slack [#rnadeseq](https://nfcore.slack.com/channels/rnadeseq) channel ([join our Slack here](https://nf-co.re/join/slack)). + +## Pipeline contribution conventions + +To make the nf-core/rnadeseq code and processing logic more understandable for new contributors and to ensure quality, we semi-standardise the way the code and other contributions are written. + +### Adding a new step + +If you wish to contribute a new step, please use the following coding standards: + +1. Define the corresponding input channel into your new process from the expected previous process channel +2. Write the process block (see below). +3. Define the output channel if needed (see below). +4. Add any new parameters to `nextflow.config` with a default (see below). +5. Add any new parameters to `nextflow_schema.json` with help text (via the `nf-core schema build` tool). +6. Add sanity checks and validation for all relevant parameters. +7. Perform local tests to validate that the new code works as expected. +8. If applicable, add a new test command in `.github/workflow/ci.yml`. +9. Update MultiQC config `assets/multiqc_config.yml` so relevant suffixes, file name clean up and module plots are in the appropriate order. If applicable, add a [MultiQC](https://https://multiqc.info/) module. +10. Add a description of the output files and if relevant any appropriate images from the MultiQC report to `docs/output.md`. + +### Default values + +Parameters should be initialised / defined with default values in `nextflow.config` under the `params` scope. + +Once there, use `nf-core schema build` to add to `nextflow_schema.json`. + +### Default processes resource requirements + +Sensible defaults for process resource requirements (CPUs / memory / time) for a process should be defined in `conf/base.config`. These should generally be specified generic with `withLabel:` selectors so they can be shared across multiple processes/steps of the pipeline. A nf-core standard set of labels that should be followed where possible can be seen in the [nf-core pipeline template](https://github.com/nf-core/tools/blob/master/nf_core/pipeline-template/conf/base.config), which has the default process as a single core-process, and then different levels of multi-core configurations for increasingly large memory requirements defined with standardised labels. + +The process resources can be passed on to the tool dynamically within the process with the `${task.cpus}` and `${task.memory}` variables in the `script:` block. + +### Naming schemes + +Please use the following naming schemes, to make it easy to understand what is going where. + +- initial process channel: `ch_output_from_` +- intermediate and terminal channels: `ch__for_` + +### Nextflow version bumping + +If you are using a new feature from core Nextflow, you may bump the minimum required version of nextflow in the pipeline with: `nf-core bump-version --nextflow . [min-nf-version]` + +### Images and figures + +For overview images and other documents we follow the nf-core [style guidelines and examples](https://nf-co.re/developers/design_guidelines). + +## GitHub Codespaces + +This repo includes a devcontainer configuration which will create a GitHub Codespaces for Nextflow development! This is an online developer environment that runs in your browser, complete with VSCode and a terminal. + +To get started: + +- Open the repo in [Codespaces](https://github.com/nf-core/rnadeseq/codespaces) +- Tools installed + - nf-core + - Nextflow + +Devcontainer specs: + +- [DevContainer config](.devcontainer/devcontainer.json) diff --git a/.github/ISSUE_TEMPLATE/bug_report.yml b/.github/ISSUE_TEMPLATE/bug_report.yml new file mode 100644 index 00000000..6e8602bd --- /dev/null +++ b/.github/ISSUE_TEMPLATE/bug_report.yml @@ -0,0 +1,50 @@ +name: Bug report +description: Report something that is broken or incorrect +labels: bug +body: + - type: markdown + attributes: + value: | + Before you post this issue, please check the documentation: + + - [nf-core website: troubleshooting](https://nf-co.re/usage/troubleshooting) + - [nf-core/rnadeseq pipeline documentation](https://nf-co.re/rnadeseq/usage) + + - type: textarea + id: description + attributes: + label: Description of the bug + description: A clear and concise description of what the bug is. + validations: + required: true + + - type: textarea + id: command_used + attributes: + label: Command used and terminal output + description: Steps to reproduce the behaviour. Please paste the command you used to launch the pipeline and the output from your terminal. + render: console + placeholder: | + $ nextflow run ... + + Some output where something broke + + - type: textarea + id: files + attributes: + label: Relevant files + description: | + Please drag and drop the relevant files here. Create a `.zip` archive if the extension is not allowed. + Your verbose log file `.nextflow.log` is often useful _(this is a hidden file in the directory where you launched the pipeline)_ as well as custom Nextflow configuration files. + + - type: textarea + id: system + attributes: + label: System information + description: | + * Nextflow version _(eg. 23.04.0)_ + * Hardware _(eg. HPC, Desktop, Cloud)_ + * Executor _(eg. slurm, local, awsbatch)_ + * Container engine: _(e.g. Docker, Singularity, Conda, Podman, Shifter, Charliecloud, or Apptainer)_ + * OS _(eg. CentOS Linux, macOS, Linux Mint)_ + * Version of nf-core/rnadeseq _(eg. 1.1, 1.5, 1.8.2)_ diff --git a/.github/ISSUE_TEMPLATE/config.yml b/.github/ISSUE_TEMPLATE/config.yml new file mode 100644 index 00000000..1c032cae --- /dev/null +++ b/.github/ISSUE_TEMPLATE/config.yml @@ -0,0 +1,7 @@ +contact_links: + - name: Join nf-core + url: https://nf-co.re/join + about: Please join the nf-core community here + - name: "Slack #rnadeseq channel" + url: https://nfcore.slack.com/channels/rnadeseq + about: Discussion about the nf-core/rnadeseq pipeline diff --git a/.github/ISSUE_TEMPLATE/feature_request.yml b/.github/ISSUE_TEMPLATE/feature_request.yml new file mode 100644 index 00000000..6bd51f6c --- /dev/null +++ b/.github/ISSUE_TEMPLATE/feature_request.yml @@ -0,0 +1,11 @@ +name: Feature request +description: Suggest an idea for the nf-core/rnadeseq pipeline +labels: enhancement +body: + - type: textarea + id: description + attributes: + label: Description of feature + description: Please describe your suggestion for a new feature. It might help to describe a problem or use case, plus any alternatives that you have considered. + validations: + required: true diff --git a/.github/PULL_REQUEST_TEMPLATE.md b/.github/PULL_REQUEST_TEMPLATE.md new file mode 100644 index 00000000..1dd64864 --- /dev/null +++ b/.github/PULL_REQUEST_TEMPLATE.md @@ -0,0 +1,26 @@ + + +## PR checklist + +- [ ] This comment contains a description of changes (with reason). +- [ ] If you've fixed a bug or added code that should be tested, add tests! +- [ ] If you've added a new tool - have you followed the pipeline conventions in the [contribution docs](https://github.com/nf-core/rnadeseq/tree/master/.github/CONTRIBUTING.md) +- [ ] If necessary, also make a PR on the nf-core/rnadeseq _branch_ on the [nf-core/test-datasets](https://github.com/nf-core/test-datasets) repository. +- [ ] Make sure your code lints (`nf-core lint`). +- [ ] Ensure the test suite passes (`nextflow run . -profile test,docker --outdir `). +- [ ] Check for unexpected warnings in debug mode (`nextflow run . -profile debug,test,docker --outdir `). +- [ ] Usage Documentation in `docs/usage.md` is updated. +- [ ] Output Documentation in `docs/output.md` is updated. +- [ ] `CHANGELOG.md` is updated. +- [ ] `README.md` is updated (including new tool citations and authors/contributors). diff --git a/.github/workflows/awsfulltest.yml b/.github/workflows/awsfulltest.yml new file mode 100644 index 00000000..8a974649 --- /dev/null +++ b/.github/workflows/awsfulltest.yml @@ -0,0 +1,39 @@ +name: nf-core AWS full size tests +# This workflow is triggered on published releases. +# It can be additionally triggered manually with GitHub actions workflow dispatch button. +# It runs the -profile 'test_full' on AWS batch + +on: + release: + types: [published] + workflow_dispatch: +jobs: + run-platform: + name: Run AWS full tests + if: github.repository == 'nf-core/rnadeseq' + runs-on: ubuntu-latest + steps: + - name: Launch workflow via Seqera Platform + uses: seqeralabs/action-tower-launch@v2 + # TODO nf-core: You can customise AWS full pipeline tests as required + # Add full size test data (but still relatively small datasets for few samples) + # on the `test_full.config` test runs with only one set of parameters + with: + workspace_id: ${{ secrets.TOWER_WORKSPACE_ID }} + access_token: ${{ secrets.TOWER_ACCESS_TOKEN }} + compute_env: ${{ secrets.TOWER_COMPUTE_ENV }} + revision: ${{ github.sha }} + workdir: s3://${{ secrets.AWS_S3_BUCKET }}/work/rnadeseq/work-${{ github.sha }} + parameters: | + { + "hook_url": "${{ secrets.MEGATESTS_ALERTS_SLACK_HOOK_URL }}", + "outdir": "s3://${{ secrets.AWS_S3_BUCKET }}/rnadeseq/results-${{ github.sha }}" + } + profiles: test_full + + - uses: actions/upload-artifact@v4 + with: + name: Seqera Platform debug log file + path: | + seqera_platform_action_*.log + seqera_platform_action_*.json diff --git a/.github/workflows/awstest.yml b/.github/workflows/awstest.yml new file mode 100644 index 00000000..b19f21e7 --- /dev/null +++ b/.github/workflows/awstest.yml @@ -0,0 +1,33 @@ +name: nf-core AWS test +# This workflow can be triggered manually with the GitHub actions workflow dispatch button. +# It runs the -profile 'test' on AWS batch + +on: + workflow_dispatch: +jobs: + run-platform: + name: Run AWS tests + if: github.repository == 'nf-core/rnadeseq' + runs-on: ubuntu-latest + steps: + # Launch workflow using Seqera Platform CLI tool action + - name: Launch workflow via Seqera Platform + uses: seqeralabs/action-tower-launch@v2 + with: + workspace_id: ${{ secrets.TOWER_WORKSPACE_ID }} + access_token: ${{ secrets.TOWER_ACCESS_TOKEN }} + compute_env: ${{ secrets.TOWER_COMPUTE_ENV }} + revision: ${{ github.sha }} + workdir: s3://${{ secrets.AWS_S3_BUCKET }}/work/rnadeseq/work-${{ github.sha }} + parameters: | + { + "outdir": "s3://${{ secrets.AWS_S3_BUCKET }}/rnadeseq/results-test-${{ github.sha }}" + } + profiles: test + + - uses: actions/upload-artifact@v4 + with: + name: Seqera Platform debug log file + path: | + seqera_platform_action_*.log + seqera_platform_action_*.json diff --git a/.github/workflows/branch.yml b/.github/workflows/branch.yml new file mode 100644 index 00000000..8cc81379 --- /dev/null +++ b/.github/workflows/branch.yml @@ -0,0 +1,44 @@ +name: nf-core branch protection +# This workflow is triggered on PRs to master branch on the repository +# It fails when someone tries to make a PR against the nf-core `master` branch instead of `dev` +on: + pull_request_target: + branches: [master] + +jobs: + test: + runs-on: ubuntu-latest + steps: + # PRs to the nf-core repo master branch are only ok if coming from the nf-core repo `dev` or any `patch` branches + - name: Check PRs + if: github.repository == 'nf-core/rnadeseq' + run: | + { [[ ${{github.event.pull_request.head.repo.full_name }} == nf-core/rnadeseq ]] && [[ $GITHUB_HEAD_REF == "dev" ]]; } || [[ $GITHUB_HEAD_REF == "patch" ]] + + # If the above check failed, post a comment on the PR explaining the failure + # NOTE - this doesn't currently work if the PR is coming from a fork, due to limitations in GitHub actions secrets + - name: Post PR comment + if: failure() + uses: mshick/add-pr-comment@b8f338c590a895d50bcbfa6c5859251edc8952fc # v2 + with: + message: | + ## This PR is against the `master` branch :x: + + * Do not close this PR + * Click _Edit_ and change the `base` to `dev` + * This CI test will remain failed until you push a new commit + + --- + + Hi @${{ github.event.pull_request.user.login }}, + + It looks like this pull-request is has been made against the [${{github.event.pull_request.head.repo.full_name }}](https://github.com/${{github.event.pull_request.head.repo.full_name }}) `master` branch. + The `master` branch on nf-core repositories should always contain code from the latest release. + Because of this, PRs to `master` are only allowed if they come from the [${{github.event.pull_request.head.repo.full_name }}](https://github.com/${{github.event.pull_request.head.repo.full_name }}) `dev` branch. + + You do not need to close this PR, you can change the target branch to `dev` by clicking the _"Edit"_ button at the top of this page. + Note that even after this, the test will continue to show as failing until you push a new commit. + + Thanks again for your contribution! + repo-token: ${{ secrets.GITHUB_TOKEN }} + allow-repeats: false diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml new file mode 100644 index 00000000..2392f9e4 --- /dev/null +++ b/.github/workflows/ci.yml @@ -0,0 +1,46 @@ +name: nf-core CI +# This workflow runs the pipeline with the minimal test dataset to check that it completes without any syntax errors +on: + push: + branches: + - dev + pull_request: + release: + types: [published] + +env: + NXF_ANSI_LOG: false + +concurrency: + group: "${{ github.workflow }}-${{ github.event.pull_request.number || github.ref }}" + cancel-in-progress: true + +jobs: + test: + name: Run pipeline with test data + # Only run on push if this is the nf-core dev branch (merged PRs) + if: "${{ github.event_name != 'push' || (github.event_name == 'push' && github.repository == 'nf-core/rnadeseq') }}" + runs-on: ubuntu-latest + strategy: + matrix: + NXF_VER: + - "23.04.0" + - "latest-everything" + steps: + - name: Check out pipeline code + uses: actions/checkout@0ad4b8fadaa221de15dcec353f45205ec38ea70b # v4 + + - name: Install Nextflow + uses: nf-core/setup-nextflow@v2 + with: + version: "${{ matrix.NXF_VER }}" + + - name: Disk space cleanup + uses: jlumbroso/free-disk-space@54081f138730dfa15788a46383842cd2f914a1be # v1.3.1 + + - name: Run pipeline with test data + # TODO nf-core: You can customise CI pipeline run tests as required + # For example: adding multiple test runs with different parameters + # Remember that you can parallelise this by using strategy.matrix + run: | + nextflow run ${GITHUB_WORKSPACE} -profile test,docker --outdir ./results diff --git a/.github/workflows/clean-up.yml b/.github/workflows/clean-up.yml new file mode 100644 index 00000000..0b6b1f27 --- /dev/null +++ b/.github/workflows/clean-up.yml @@ -0,0 +1,24 @@ +name: "Close user-tagged issues and PRs" +on: + schedule: + - cron: "0 0 * * 0" # Once a week + +jobs: + clean-up: + runs-on: ubuntu-latest + permissions: + issues: write + pull-requests: write + steps: + - uses: actions/stale@28ca1036281a5e5922ead5184a1bbf96e5fc984e # v9 + with: + stale-issue-message: "This issue has been tagged as awaiting-changes or awaiting-feedback by an nf-core contributor. Remove stale label or add a comment otherwise this issue will be closed in 20 days." + stale-pr-message: "This PR has been tagged as awaiting-changes or awaiting-feedback by an nf-core contributor. Remove stale label or add a comment if it is still useful." + close-issue-message: "This issue was closed because it has been tagged as awaiting-changes or awaiting-feedback by an nf-core contributor and then staled for 20 days with no activity." + days-before-stale: 30 + days-before-close: 20 + days-before-pr-close: -1 + any-of-labels: "awaiting-changes,awaiting-feedback" + exempt-issue-labels: "WIP" + exempt-pr-labels: "WIP" + repo-token: "${{ secrets.GITHUB_TOKEN }}" diff --git a/.github/workflows/download_pipeline.yml b/.github/workflows/download_pipeline.yml new file mode 100644 index 00000000..2d20d644 --- /dev/null +++ b/.github/workflows/download_pipeline.yml @@ -0,0 +1,86 @@ +name: Test successful pipeline download with 'nf-core download' + +# Run the workflow when: +# - dispatched manually +# - when a PR is opened or reopened to master branch +# - the head branch of the pull request is updated, i.e. if fixes for a release are pushed last minute to dev. +on: + workflow_dispatch: + inputs: + testbranch: + description: "The specific branch you wish to utilize for the test execution of nf-core download." + required: true + default: "dev" + pull_request: + types: + - opened + - edited + - synchronize + branches: + - master + pull_request_target: + branches: + - master + +env: + NXF_ANSI_LOG: false + +jobs: + download: + runs-on: ubuntu-latest + steps: + - name: Install Nextflow + uses: nf-core/setup-nextflow@v2 + + - name: Disk space cleanup + uses: jlumbroso/free-disk-space@54081f138730dfa15788a46383842cd2f914a1be # v1.3.1 + + - uses: actions/setup-python@82c7e631bb3cdc910f68e0081d67478d79c6982d # v5 + with: + python-version: "3.12" + architecture: "x64" + - uses: eWaterCycle/setup-singularity@931d4e31109e875b13309ae1d07c70ca8fbc8537 # v7 + with: + singularity-version: 3.8.3 + + - name: Install dependencies + run: | + python -m pip install --upgrade pip + pip install git+https://github.com/nf-core/tools.git@dev + + - name: Get the repository name and current branch set as environment variable + run: | + echo "REPO_LOWERCASE=${GITHUB_REPOSITORY,,}" >> ${GITHUB_ENV} + echo "REPOTITLE_LOWERCASE=$(basename ${GITHUB_REPOSITORY,,})" >> ${GITHUB_ENV} + echo "REPO_BRANCH=${{ github.event.inputs.testbranch || 'dev' }}" >> ${GITHUB_ENV} + + - name: Download the pipeline + env: + NXF_SINGULARITY_CACHEDIR: ./ + run: | + nf-core download ${{ env.REPO_LOWERCASE }} \ + --revision ${{ env.REPO_BRANCH }} \ + --outdir ./${{ env.REPOTITLE_LOWERCASE }} \ + --compress "none" \ + --container-system 'singularity' \ + --container-library "quay.io" -l "docker.io" -l "ghcr.io" \ + --container-cache-utilisation 'amend' \ + --download-configuration + + - name: Inspect download + run: tree ./${{ env.REPOTITLE_LOWERCASE }} + + - name: Run the downloaded pipeline (stub) + id: stub_run_pipeline + continue-on-error: true + env: + NXF_SINGULARITY_CACHEDIR: ./ + NXF_SINGULARITY_HOME_MOUNT: true + run: nextflow run ./${{ env.REPOTITLE_LOWERCASE }}/$( sed 's/\W/_/g' <<< ${{ env.REPO_BRANCH }}) -stub -profile test,singularity --outdir ./results + - name: Run the downloaded pipeline (stub run not supported) + id: run_pipeline + if: ${{ job.steps.stub_run_pipeline.status == failure() }} + env: + NXF_SINGULARITY_CACHEDIR: ./ + NXF_SINGULARITY_HOME_MOUNT: true + run: nextflow run ./${{ env.REPOTITLE_LOWERCASE }}/$( sed 's/\W/_/g' <<< ${{ env.REPO_BRANCH }}) -profile test,singularity --outdir ./results diff --git a/.github/workflows/fix-linting.yml b/.github/workflows/fix-linting.yml new file mode 100644 index 00000000..d0076123 --- /dev/null +++ b/.github/workflows/fix-linting.yml @@ -0,0 +1,89 @@ +name: Fix linting from a comment +on: + issue_comment: + types: [created] + +jobs: + fix-linting: + # Only run if comment is on a PR with the main repo, and if it contains the magic keywords + if: > + contains(github.event.comment.html_url, '/pull/') && + contains(github.event.comment.body, '@nf-core-bot fix linting') && + github.repository == 'nf-core/rnadeseq' + runs-on: ubuntu-latest + steps: + # Use the @nf-core-bot token to check out so we can push later + - uses: actions/checkout@0ad4b8fadaa221de15dcec353f45205ec38ea70b # v4 + with: + token: ${{ secrets.nf_core_bot_auth_token }} + + # indication that the linting is being fixed + - name: React on comment + uses: peter-evans/create-or-update-comment@71345be0265236311c031f5c7866368bd1eff043 # v4 + with: + comment-id: ${{ github.event.comment.id }} + reactions: eyes + + # Action runs on the issue comment, so we don't get the PR by default + # Use the gh cli to check out the PR + - name: Checkout Pull Request + run: gh pr checkout ${{ github.event.issue.number }} + env: + GITHUB_TOKEN: ${{ secrets.nf_core_bot_auth_token }} + + # Install and run pre-commit + - uses: actions/setup-python@82c7e631bb3cdc910f68e0081d67478d79c6982d # v5 + with: + python-version: "3.12" + + - name: Install pre-commit + run: pip install pre-commit + + - name: Run pre-commit + id: pre-commit + run: pre-commit run --all-files + continue-on-error: true + + # indication that the linting has finished + - name: react if linting finished succesfully + if: steps.pre-commit.outcome == 'success' + uses: peter-evans/create-or-update-comment@71345be0265236311c031f5c7866368bd1eff043 # v4 + with: + comment-id: ${{ github.event.comment.id }} + reactions: "+1" + + - name: Commit & push changes + id: commit-and-push + if: steps.pre-commit.outcome == 'failure' + run: | + git config user.email "core@nf-co.re" + git config user.name "nf-core-bot" + git config push.default upstream + git add . + git status + git commit -m "[automated] Fix code linting" + git push + + - name: react if linting errors were fixed + id: react-if-fixed + if: steps.commit-and-push.outcome == 'success' + uses: peter-evans/create-or-update-comment@71345be0265236311c031f5c7866368bd1eff043 # v4 + with: + comment-id: ${{ github.event.comment.id }} + reactions: hooray + + - name: react if linting errors were not fixed + if: steps.commit-and-push.outcome == 'failure' + uses: peter-evans/create-or-update-comment@71345be0265236311c031f5c7866368bd1eff043 # v4 + with: + comment-id: ${{ github.event.comment.id }} + reactions: confused + + - name: react if linting errors were not fixed + if: steps.commit-and-push.outcome == 'failure' + uses: peter-evans/create-or-update-comment@71345be0265236311c031f5c7866368bd1eff043 # v4 + with: + issue-number: ${{ github.event.issue.number }} + body: | + @${{ github.actor }} I tried to fix the linting errors, but it didn't work. Please fix them manually. + See [CI log](https://github.com/nf-core/rnadeseq/actions/runs/${{ github.run_id }}) for more details. diff --git a/.github/workflows/linting.yml b/.github/workflows/linting.yml new file mode 100644 index 00000000..1fcafe88 --- /dev/null +++ b/.github/workflows/linting.yml @@ -0,0 +1,68 @@ +name: nf-core linting +# This workflow is triggered on pushes and PRs to the repository. +# It runs the `nf-core lint` and markdown lint tests to ensure +# that the code meets the nf-core guidelines. +on: + push: + branches: + - dev + pull_request: + release: + types: [published] + +jobs: + pre-commit: + runs-on: ubuntu-latest + steps: + - uses: actions/checkout@0ad4b8fadaa221de15dcec353f45205ec38ea70b # v4 + + - name: Set up Python 3.12 + uses: actions/setup-python@82c7e631bb3cdc910f68e0081d67478d79c6982d # v5 + with: + python-version: "3.12" + + - name: Install pre-commit + run: pip install pre-commit + + - name: Run pre-commit + run: pre-commit run --all-files + + nf-core: + runs-on: ubuntu-latest + steps: + - name: Check out pipeline code + uses: actions/checkout@0ad4b8fadaa221de15dcec353f45205ec38ea70b # v4 + + - name: Install Nextflow + uses: nf-core/setup-nextflow@v2 + + - uses: actions/setup-python@82c7e631bb3cdc910f68e0081d67478d79c6982d # v5 + with: + python-version: "3.12" + architecture: "x64" + + - name: Install dependencies + run: | + python -m pip install --upgrade pip + pip install nf-core + + - name: Run nf-core lint + env: + GITHUB_COMMENTS_URL: ${{ github.event.pull_request.comments_url }} + GITHUB_TOKEN: ${{ secrets.GITHUB_TOKEN }} + GITHUB_PR_COMMIT: ${{ github.event.pull_request.head.sha }} + run: nf-core -l lint_log.txt lint --dir ${GITHUB_WORKSPACE} --markdown lint_results.md + + - name: Save PR number + if: ${{ always() }} + run: echo ${{ github.event.pull_request.number }} > PR_number.txt + + - name: Upload linting log file artifact + if: ${{ always() }} + uses: actions/upload-artifact@65462800fd760344b1a7b4382951275a0abb4808 # v4 + with: + name: linting-logs + path: | + lint_log.txt + lint_results.md + PR_number.txt diff --git a/.github/workflows/linting_comment.yml b/.github/workflows/linting_comment.yml new file mode 100644 index 00000000..40acc23f --- /dev/null +++ b/.github/workflows/linting_comment.yml @@ -0,0 +1,28 @@ +name: nf-core linting comment +# This workflow is triggered after the linting action is complete +# It posts an automated comment to the PR, even if the PR is coming from a fork + +on: + workflow_run: + workflows: ["nf-core linting"] + +jobs: + test: + runs-on: ubuntu-latest + steps: + - name: Download lint results + uses: dawidd6/action-download-artifact@09f2f74827fd3a8607589e5ad7f9398816f540fe # v3 + with: + workflow: linting.yml + workflow_conclusion: completed + + - name: Get PR number + id: pr_number + run: echo "pr_number=$(cat linting-logs/PR_number.txt)" >> $GITHUB_OUTPUT + + - name: Post PR comment + uses: marocchino/sticky-pull-request-comment@331f8f5b4215f0445d3c07b4967662a32a2d3e31 # v2 + with: + GITHUB_TOKEN: ${{ secrets.GITHUB_TOKEN }} + number: ${{ steps.pr_number.outputs.pr_number }} + path: linting-logs/lint_results.md diff --git a/.github/workflows/release-announcements.yml b/.github/workflows/release-announcements.yml new file mode 100644 index 00000000..03ecfcf7 --- /dev/null +++ b/.github/workflows/release-announcements.yml @@ -0,0 +1,75 @@ +name: release-announcements +# Automatic release toot and tweet anouncements +on: + release: + types: [published] + workflow_dispatch: + +jobs: + toot: + runs-on: ubuntu-latest + steps: + - name: get topics and convert to hashtags + id: get_topics + run: | + echo "topics=$(curl -s https://nf-co.re/pipelines.json | jq -r '.remote_workflows[] | select(.full_name == "${{ github.repository }}") | .topics[]' | awk '{print "#"$0}' | tr '\n' ' ')" >> $GITHUB_OUTPUT + + - uses: rzr/fediverse-action@master + with: + access-token: ${{ secrets.MASTODON_ACCESS_TOKEN }} + host: "mstdn.science" # custom host if not "mastodon.social" (default) + # GitHub event payload + # https://docs.github.com/en/developers/webhooks-and-events/webhooks/webhook-events-and-payloads#release + message: | + Pipeline release! ${{ github.repository }} v${{ github.event.release.tag_name }} - ${{ github.event.release.name }}! + + Please see the changelog: ${{ github.event.release.html_url }} + + ${{ steps.get_topics.outputs.topics }} #nfcore #openscience #nextflow #bioinformatics + + send-tweet: + runs-on: ubuntu-latest + + steps: + - uses: actions/setup-python@82c7e631bb3cdc910f68e0081d67478d79c6982d # v5 + with: + python-version: "3.10" + - name: Install dependencies + run: pip install tweepy==4.14.0 + - name: Send tweet + shell: python + run: | + import os + import tweepy + + client = tweepy.Client( + access_token=os.getenv("TWITTER_ACCESS_TOKEN"), + access_token_secret=os.getenv("TWITTER_ACCESS_TOKEN_SECRET"), + consumer_key=os.getenv("TWITTER_CONSUMER_KEY"), + consumer_secret=os.getenv("TWITTER_CONSUMER_SECRET"), + ) + tweet = os.getenv("TWEET") + client.create_tweet(text=tweet) + env: + TWEET: | + Pipeline release! ${{ github.repository }} v${{ github.event.release.tag_name }} - ${{ github.event.release.name }}! + + Please see the changelog: ${{ github.event.release.html_url }} + TWITTER_CONSUMER_KEY: ${{ secrets.TWITTER_CONSUMER_KEY }} + TWITTER_CONSUMER_SECRET: ${{ secrets.TWITTER_CONSUMER_SECRET }} + TWITTER_ACCESS_TOKEN: ${{ secrets.TWITTER_ACCESS_TOKEN }} + TWITTER_ACCESS_TOKEN_SECRET: ${{ secrets.TWITTER_ACCESS_TOKEN_SECRET }} + + bsky-post: + runs-on: ubuntu-latest + steps: + - uses: zentered/bluesky-post-action@80dbe0a7697de18c15ad22f4619919ceb5ccf597 # v0.1.0 + with: + post: | + Pipeline release! ${{ github.repository }} v${{ github.event.release.tag_name }} - ${{ github.event.release.name }}! + + Please see the changelog: ${{ github.event.release.html_url }} + env: + BSKY_IDENTIFIER: ${{ secrets.BSKY_IDENTIFIER }} + BSKY_PASSWORD: ${{ secrets.BSKY_PASSWORD }} + # diff --git a/.gitignore b/.gitignore new file mode 100644 index 00000000..5124c9ac --- /dev/null +++ b/.gitignore @@ -0,0 +1,8 @@ +.nextflow* +work/ +data/ +results/ +.DS_Store +testing/ +testing* +*.pyc diff --git a/.gitpod.yml b/.gitpod.yml new file mode 100644 index 00000000..105a1821 --- /dev/null +++ b/.gitpod.yml @@ -0,0 +1,20 @@ +image: nfcore/gitpod:latest +tasks: + - name: Update Nextflow and setup pre-commit + command: | + pre-commit install --install-hooks + nextflow self-update + - name: unset JAVA_TOOL_OPTIONS + command: | + unset JAVA_TOOL_OPTIONS + +vscode: + extensions: # based on nf-core.nf-core-extensionpack + - esbenp.prettier-vscode # Markdown/CommonMark linting and style checking for Visual Studio Code + - EditorConfig.EditorConfig # override user/workspace settings with settings found in .editorconfig files + - Gruntfuggly.todo-tree # Display TODO and FIXME in a tree view in the activity bar + - mechatroner.rainbow-csv # Highlight columns in csv files in different colors + # - nextflow.nextflow # Nextflow syntax highlighting + - oderwat.indent-rainbow # Highlight indentation level + - streetsidesoftware.code-spell-checker # Spelling checker for source code + - charliermarsh.ruff # Code linter Ruff diff --git a/.nf-core.yml b/.nf-core.yml new file mode 100644 index 00000000..e0b85a77 --- /dev/null +++ b/.nf-core.yml @@ -0,0 +1,2 @@ +repository_type: pipeline +nf_core_version: "2.14.1" diff --git a/.pre-commit-config.yaml b/.pre-commit-config.yaml new file mode 100644 index 00000000..4dc0f1dc --- /dev/null +++ b/.pre-commit-config.yaml @@ -0,0 +1,13 @@ +repos: + - repo: https://github.com/pre-commit/mirrors-prettier + rev: "v3.1.0" + hooks: + - id: prettier + additional_dependencies: + - prettier@3.2.5 + + - repo: https://github.com/editorconfig-checker/editorconfig-checker.python + rev: "2.7.3" + hooks: + - id: editorconfig-checker + alias: ec diff --git a/.prettierignore b/.prettierignore new file mode 100644 index 00000000..437d763d --- /dev/null +++ b/.prettierignore @@ -0,0 +1,12 @@ +email_template.html +adaptivecard.json +slackreport.json +.nextflow* +work/ +data/ +results/ +.DS_Store +testing/ +testing* +*.pyc +bin/ diff --git a/.prettierrc.yml b/.prettierrc.yml new file mode 100644 index 00000000..c81f9a76 --- /dev/null +++ b/.prettierrc.yml @@ -0,0 +1 @@ +printWidth: 120 diff --git a/CHANGELOG.md b/CHANGELOG.md new file mode 100644 index 00000000..4c00d15a --- /dev/null +++ b/CHANGELOG.md @@ -0,0 +1,16 @@ +# nf-core/rnadeseq: Changelog + +The format is based on [Keep a Changelog](https://keepachangelog.com/en/1.0.0/) +and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0.html). + +## v1.0dev - [date] + +Initial release of nf-core/rnadeseq, created with the [nf-core](https://nf-co.re/) template. + +### `Added` + +### `Fixed` + +### `Dependencies` + +### `Deprecated` diff --git a/CITATIONS.md b/CITATIONS.md new file mode 100644 index 00000000..56ec302a --- /dev/null +++ b/CITATIONS.md @@ -0,0 +1,41 @@ +# nf-core/rnadeseq: Citations + +## [nf-core](https://pubmed.ncbi.nlm.nih.gov/32055031/) + +> Ewels PA, Peltzer A, Fillinger S, Patel H, Alneberg J, Wilm A, Garcia MU, Di Tommaso P, Nahnsen S. The nf-core framework for community-curated bioinformatics pipelines. Nat Biotechnol. 2020 Mar;38(3):276-278. doi: 10.1038/s41587-020-0439-x. PubMed PMID: 32055031. + +## [Nextflow](https://pubmed.ncbi.nlm.nih.gov/28398311/) + +> Di Tommaso P, Chatzou M, Floden EW, Barja PP, Palumbo E, Notredame C. Nextflow enables reproducible computational workflows. Nat Biotechnol. 2017 Apr 11;35(4):316-319. doi: 10.1038/nbt.3820. PubMed PMID: 28398311. + +## Pipeline tools + +- [FastQC](https://www.bioinformatics.babraham.ac.uk/projects/fastqc/) + + > Andrews, S. (2010). FastQC: A Quality Control Tool for High Throughput Sequence Data [Online]. + +- [MultiQC](https://pubmed.ncbi.nlm.nih.gov/27312411/) + + > Ewels P, Magnusson M, Lundin S, Käller M. MultiQC: summarize analysis results for multiple tools and samples in a single report. Bioinformatics. 2016 Oct 1;32(19):3047-8. doi: 10.1093/bioinformatics/btw354. Epub 2016 Jun 16. PubMed PMID: 27312411; PubMed Central PMCID: PMC5039924. + +## Software packaging/containerisation tools + +- [Anaconda](https://anaconda.com) + + > Anaconda Software Distribution. Computer software. Vers. 2-2.4.0. Anaconda, Nov. 2016. Web. + +- [Bioconda](https://pubmed.ncbi.nlm.nih.gov/29967506/) + + > Grüning B, Dale R, Sjödin A, Chapman BA, Rowe J, Tomkins-Tinch CH, Valieris R, Köster J; Bioconda Team. Bioconda: sustainable and comprehensive software distribution for the life sciences. Nat Methods. 2018 Jul;15(7):475-476. doi: 10.1038/s41592-018-0046-7. PubMed PMID: 29967506. + +- [BioContainers](https://pubmed.ncbi.nlm.nih.gov/28379341/) + + > da Veiga Leprevost F, Grüning B, Aflitos SA, Röst HL, Uszkoreit J, Barsnes H, Vaudel M, Moreno P, Gatto L, Weber J, Bai M, Jimenez RC, Sachsenberg T, Pfeuffer J, Alvarez RV, Griss J, Nesvizhskii AI, Perez-Riverol Y. BioContainers: an open-source and community-driven framework for software standardization. Bioinformatics. 2017 Aug 15;33(16):2580-2582. doi: 10.1093/bioinformatics/btx192. PubMed PMID: 28379341; PubMed Central PMCID: PMC5870671. + +- [Docker](https://dl.acm.org/doi/10.5555/2600239.2600241) + + > Merkel, D. (2014). Docker: lightweight linux containers for consistent development and deployment. Linux Journal, 2014(239), 2. doi: 10.5555/2600239.2600241. + +- [Singularity](https://pubmed.ncbi.nlm.nih.gov/28494014/) + + > Kurtzer GM, Sochat V, Bauer MW. Singularity: Scientific containers for mobility of compute. PLoS One. 2017 May 11;12(5):e0177459. doi: 10.1371/journal.pone.0177459. eCollection 2017. PubMed PMID: 28494014; PubMed Central PMCID: PMC5426675. diff --git a/CODE_OF_CONDUCT.md b/CODE_OF_CONDUCT.md new file mode 100644 index 00000000..c089ec78 --- /dev/null +++ b/CODE_OF_CONDUCT.md @@ -0,0 +1,182 @@ +# Code of Conduct at nf-core (v1.4) + +## Our Pledge + +In the interest of fostering an open, collaborative, and welcoming environment, we as contributors and maintainers of nf-core pledge to making participation in our projects and community a harassment-free experience for everyone, regardless of: + +- Age +- Ability +- Body size +- Caste +- Familial status +- Gender identity and expression +- Geographical location +- Level of experience +- Nationality and national origins +- Native language +- Neurodiversity +- Race or ethnicity +- Religion +- Sexual identity and orientation +- Socioeconomic status + +Please note that the list above is alphabetised and is therefore not ranked in any order of preference or importance. + +## Preamble + +:::note +This Code of Conduct (CoC) has been drafted by Renuka Kudva, Cris Tuñí, and Michael Heuer, with input from the nf-core Core Team and Susanna Marquez from the nf-core community. "We", in this document, refers to the Safety Officers and members of the nf-core Core Team, both of whom are deemed to be members of the nf-core community and are therefore required to abide by this Code of Conduct. This document will be amended periodically to keep it up-to-date. In case of any dispute, the most current version will apply. +::: + +An up-to-date list of members of the nf-core core team can be found [here](https://nf-co.re/about). + +Our Safety Officers are Saba Nafees, Cris Tuñí, and Michael Heuer. + +nf-core is a young and growing community that welcomes contributions from anyone with a shared vision for [Open Science Policies](https://www.fosteropenscience.eu/taxonomy/term/8). Open science policies encompass inclusive behaviours and we strive to build and maintain a safe and inclusive environment for all individuals. + +We have therefore adopted this CoC, which we require all members of our community and attendees of nf-core events to adhere to in all our workspaces at all times. Workspaces include, but are not limited to, Slack, meetings on Zoom, gather.town, YouTube live etc. + +Our CoC will be strictly enforced and the nf-core team reserves the right to exclude participants who do not comply with our guidelines from our workspaces and future nf-core activities. + +We ask all members of our community to help maintain supportive and productive workspaces and to avoid behaviours that can make individuals feel unsafe or unwelcome. Please help us maintain and uphold this CoC. + +Questions, concerns, or ideas on what we can include? Contact members of the Safety Team on Slack or email safety [at] nf-co [dot] re. + +## Our Responsibilities + +Members of the Safety Team (the Safety Officers) are responsible for clarifying the standards of acceptable behavior and are expected to take appropriate and fair corrective action in response to any instances of unacceptable behaviour. + +The Safety Team, in consultation with the nf-core core team, have the right and responsibility to remove, edit, or reject comments, commits, code, wiki edits, issues, and other contributions that are not aligned to this CoC, or to ban temporarily or permanently any contributor for other behaviors that they deem inappropriate, threatening, offensive, or harmful. + +Members of the core team or the Safety Team who violate the CoC will be required to recuse themselves pending investigation. They will not have access to any reports of the violations and will be subject to the same actions as others in violation of the CoC. + +## When and where does this Code of Conduct apply? + +Participation in the nf-core community is contingent on following these guidelines in all our workspaces and events, such as hackathons, workshops, bytesize, and collaborative workspaces on gather.town. These guidelines include, but are not limited to, the following (listed alphabetically and therefore in no order of preference): + +- Communicating with an official project email address. +- Communicating with community members within the nf-core Slack channel. +- Participating in hackathons organised by nf-core (both online and in-person events). +- Participating in collaborative work on GitHub, Google Suite, community calls, mentorship meetings, email correspondence, and on the nf-core gather.town workspace. +- Participating in workshops, training, and seminar series organised by nf-core (both online and in-person events). This applies to events hosted on web-based platforms such as Zoom, gather.town, Jitsi, YouTube live etc. +- Representing nf-core on social media. This includes both official and personal accounts. + +## nf-core cares 😊 + +nf-core's CoC and expectations of respectful behaviours for all participants (including organisers and the nf-core team) include, but are not limited to, the following (listed in alphabetical order): + +- Ask for consent before sharing another community member’s personal information (including photographs) on social media. +- Be respectful of differing viewpoints and experiences. We are all here to learn from one another and a difference in opinion can present a good learning opportunity. +- Celebrate your accomplishments! (Get creative with your use of emojis 🎉 🥳 💯 🙌 !) +- Demonstrate empathy towards other community members. (We don’t all have the same amount of time to dedicate to nf-core. If tasks are pending, don’t hesitate to gently remind members of your team. If you are leading a task, ask for help if you feel overwhelmed.) +- Engage with and enquire after others. (This is especially important given the geographically remote nature of the nf-core community, so let’s do this the best we can) +- Focus on what is best for the team and the community. (When in doubt, ask) +- Accept feedback, yet be unafraid to question, deliberate, and learn. +- Introduce yourself to members of the community. (We’ve all been outsiders and we know that talking to strangers can be hard for some, but remember we’re interested in getting to know you and your visions for open science!) +- Show appreciation and **provide clear feedback**. (This is especially important because we don’t see each other in person and it can be harder to interpret subtleties. Also remember that not everyone understands a certain language to the same extent as you do, so **be clear in your communication to be kind.**) +- Take breaks when you feel like you need them. +- Use welcoming and inclusive language. (Participants are encouraged to display their chosen pronouns on Zoom or in communication on Slack) + +## nf-core frowns on 😕 + +The following behaviours from any participants within the nf-core community (including the organisers) will be considered unacceptable under this CoC. Engaging or advocating for any of the following could result in expulsion from nf-core workspaces: + +- Deliberate intimidation, stalking or following and sustained disruption of communication among participants of the community. This includes hijacking shared screens through actions such as using the annotate tool in conferencing software such as Zoom. +- “Doxing” i.e. posting (or threatening to post) another person’s personal identifying information online. +- Spamming or trolling of individuals on social media. +- Use of sexual or discriminatory imagery, comments, jokes, or unwelcome sexual attention. +- Verbal and text comments that reinforce social structures of domination related to gender, gender identity and expression, sexual orientation, ability, physical appearance, body size, race, age, religion, or work experience. + +### Online Trolling + +The majority of nf-core interactions and events are held online. Unfortunately, holding events online comes with the risk of online trolling. This is unacceptable — reports of such behaviour will be taken very seriously and perpetrators will be excluded from activities immediately. + +All community members are **required** to ask members of the group they are working with for explicit consent prior to taking screenshots of individuals during video calls. + +## Procedures for reporting CoC violations + +If someone makes you feel uncomfortable through their behaviours or actions, report it as soon as possible. + +You can reach out to members of the Safety Team (Saba Nafees, Cris Tuñí, and Michael Heuer) on Slack. Alternatively, contact a member of the nf-core core team [nf-core core team](https://nf-co.re/about), and they will forward your concerns to the Safety Team. + +Issues directly concerning members of the Core Team or the Safety Team will be dealt with by other members of the core team and the safety manager — possible conflicts of interest will be taken into account. nf-core is also in discussions about having an ombudsperson and details will be shared in due course. + +All reports will be handled with the utmost discretion and confidentiality. + +You can also report any CoC violations to safety [at] nf-co [dot] re. In your email report, please do your best to include: + +- Your contact information. +- Identifying information (e.g. names, nicknames, pseudonyms) of the participant who has violated the Code of Conduct. +- The behaviour that was in violation and the circumstances surrounding the incident. +- The approximate time of the behaviour (if different than the time the report was made). +- Other people involved in the incident, if applicable. +- If you believe the incident is ongoing. +- If there is a publicly available record (e.g. mailing list record, a screenshot). +- Any additional information. + +After you file a report, one or more members of our Safety Team will contact you to follow up on your report. + +## Who will read and handle reports + +All reports will be read and handled by the members of the Safety Team at nf-core. + +If members of the Safety Team are deemed to have a conflict of interest with a report, they will be required to recuse themselves as per our Code of Conduct and will not have access to any follow-ups. + +To keep this first report confidential from any of the Safety Team members, please submit your first report by direct messaging on Slack/direct email to any of the nf-core members you are comfortable disclosing the information to, and be explicit about which member(s) you do not consent to sharing the information with. + +## Reviewing reports + +After receiving the report, members of the Safety Team will review the incident report to determine whether immediate action is required, for example, whether there is immediate threat to participants’ safety. + +The Safety Team, in consultation with members of the nf-core core team, will assess the information to determine whether the report constitutes a Code of Conduct violation, for them to decide on a course of action. + +In the case of insufficient information, one or more members of the Safety Team may contact the reporter, the reportee, or any other attendees to obtain more information. + +Once additional information is gathered, the Safety Team will collectively review and decide on the best course of action to take, if any. The Safety Team reserves the right to not act on a report. + +## Confidentiality + +All reports, and any additional information included, are only shared with the team of safety officers (and possibly members of the core team, in case the safety officer is in violation of the CoC). We will respect confidentiality requests for the purpose of protecting victims of abuse. + +We will not name harassment victims, beyond discussions between the safety officer and members of the nf-core team, without the explicit consent of the individuals involved. + +## Enforcement + +Actions taken by the nf-core’s Safety Team may include, but are not limited to: + +- Asking anyone to stop a behaviour. +- Asking anyone to leave the event and online spaces either temporarily, for the remainder of the event, or permanently. +- Removing access to the gather.town and Slack, either temporarily or permanently. +- Communicating to all participants to reinforce our expectations for conduct and remind what is unacceptable behaviour; this may be public for practical reasons. +- Communicating to all participants that an incident has taken place and how we will act or have acted — this may be for the purpose of letting event participants know we are aware of and dealing with the incident. +- Banning anyone from participating in nf-core-managed spaces, future events, and activities, either temporarily or permanently. +- No action. + +## Attribution and Acknowledgements + +- The [Contributor Covenant, version 1.4](http://contributor-covenant.org/version/1/4) +- The [OpenCon 2017 Code of Conduct](http://www.opencon2017.org/code_of_conduct) (CC BY 4.0 OpenCon organisers, SPARC and Right to Research Coalition) +- The [eLife innovation sprint 2020 Code of Conduct](https://sprint.elifesciences.org/code-of-conduct/) +- The [Mozilla Community Participation Guidelines v3.1](https://www.mozilla.org/en-US/about/governance/policies/participation/) (version 3.1, CC BY-SA 3.0 Mozilla) + +## Changelog + +### v1.4 - February 8th, 2022 + +- Included a new member of the Safety Team. Corrected a typographical error in the text. + +### v1.3 - December 10th, 2021 + +- Added a statement that the CoC applies to nf-core gather.town workspaces. Corrected typographical errors in the text. + +### v1.2 - November 12th, 2021 + +- Removed information specific to reporting CoC violations at the Hackathon in October 2021. + +### v1.1 - October 14th, 2021 + +- Updated with names of new Safety Officers and specific information for the hackathon in October 2021. + +### v1.0 - March 15th, 2021 + +- Complete rewrite from original [Contributor Covenant](http://contributor-covenant.org/) CoC. diff --git a/LICENSE b/LICENSE new file mode 100644 index 00000000..b0c71392 --- /dev/null +++ b/LICENSE @@ -0,0 +1,21 @@ +MIT License + +Copyright (c) + +Permission is hereby granted, free of charge, to any person obtaining a copy +of this software and associated documentation files (the "Software"), to deal +in the Software without restriction, including without limitation the rights +to use, copy, modify, merge, publish, distribute, sublicense, and/or sell +copies of the Software, and to permit persons to whom the Software is +furnished to do so, subject to the following conditions: + +The above copyright notice and this permission notice shall be included in all +copies or substantial portions of the Software. + +THE SOFTWARE IS PROVIDED "AS IS", WITHOUT WARRANTY OF ANY KIND, EXPRESS OR +IMPLIED, INCLUDING BUT NOT LIMITED TO THE WARRANTIES OF MERCHANTABILITY, +FITNESS FOR A PARTICULAR PURPOSE AND NONINFRINGEMENT. IN NO EVENT SHALL THE +AUTHORS OR COPYRIGHT HOLDERS BE LIABLE FOR ANY CLAIM, DAMAGES OR OTHER +LIABILITY, WHETHER IN AN ACTION OF CONTRACT, TORT OR OTHERWISE, ARISING FROM, +OUT OF OR IN CONNECTION WITH THE SOFTWARE OR THE USE OR OTHER DEALINGS IN THE +SOFTWARE. diff --git a/README.md b/README.md new file mode 100644 index 00000000..5f6a700e --- /dev/null +++ b/README.md @@ -0,0 +1,110 @@ +

+ + + nf-core/rnadeseq + +

+ +[![GitHub Actions CI Status](https://github.com/nf-core/rnadeseq/actions/workflows/ci.yml/badge.svg)](https://github.com/nf-core/rnadeseq/actions/workflows/ci.yml) +[![GitHub Actions Linting Status](https://github.com/nf-core/rnadeseq/actions/workflows/linting.yml/badge.svg)](https://github.com/nf-core/rnadeseq/actions/workflows/linting.yml)[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/rnadeseq/results)[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.XXXXXXX-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.XXXXXXX) +[![nf-test](https://img.shields.io/badge/unit_tests-nf--test-337ab7.svg)](https://www.nf-test.com) + +[![Nextflow](https://img.shields.io/badge/nextflow%20DSL2-%E2%89%A523.04.0-23aa62.svg)](https://www.nextflow.io/) +[![run with conda](http://img.shields.io/badge/run%20with-conda-3EB049?labelColor=000000&logo=anaconda)](https://docs.conda.io/en/latest/) +[![run with docker](https://img.shields.io/badge/run%20with-docker-0db7ed?labelColor=000000&logo=docker)](https://www.docker.com/) +[![run with singularity](https://img.shields.io/badge/run%20with-singularity-1d355c.svg?labelColor=000000)](https://sylabs.io/docs/) +[![Launch on Seqera Platform](https://img.shields.io/badge/Launch%20%F0%9F%9A%80-Seqera%20Platform-%234256e7)](https://cloud.seqera.io/launch?pipeline=https://github.com/nf-core/rnadeseq) + +[![Get help on Slack](http://img.shields.io/badge/slack-nf--core%20%23rnadeseq-4A154B?labelColor=000000&logo=slack)](https://nfcore.slack.com/channels/rnadeseq)[![Follow on Twitter](http://img.shields.io/badge/twitter-%40nf__core-1DA1F2?labelColor=000000&logo=twitter)](https://twitter.com/nf_core)[![Follow on Mastodon](https://img.shields.io/badge/mastodon-nf__core-6364ff?labelColor=FFFFFF&logo=mastodon)](https://mstdn.science/@nf_core)[![Watch on YouTube](http://img.shields.io/badge/youtube-nf--core-FF0000?labelColor=000000&logo=youtube)](https://www.youtube.com/c/nf-core) + +## Introduction + +**nf-core/rnadeseq** is a bioinformatics pipeline that ... + + + + + + +1. Read QC ([`FastQC`](https://www.bioinformatics.babraham.ac.uk/projects/fastqc/)) +2. Present QC for raw reads ([`MultiQC`](http://multiqc.info/)) + +## Usage + +> [!NOTE] +> If you are new to Nextflow and nf-core, please refer to [this page](https://nf-co.re/docs/usage/installation) on how to set-up Nextflow. Make sure to [test your setup](https://nf-co.re/docs/usage/introduction#how-to-run-a-pipeline) with `-profile test` before running the workflow on actual data. + + + +Now, you can run the pipeline using: + + + +```bash +nextflow run nf-core/rnadeseq \ + -profile \ + --input samplesheet.csv \ + --outdir +``` + +> [!WARNING] +> Please provide pipeline parameters via the CLI or Nextflow `-params-file` option. Custom config files including those provided by the `-c` Nextflow option can be used to provide any configuration _**except for parameters**_; +> see [docs](https://nf-co.re/usage/configuration#custom-configuration-files). + +For more details and further functionality, please refer to the [usage documentation](https://nf-co.re/rnadeseq/usage) and the [parameter documentation](https://nf-co.re/rnadeseq/parameters). + +## Pipeline output + +To see the results of an example test run with a full size dataset refer to the [results](https://nf-co.re/rnadeseq/results) tab on the nf-core website pipeline page. +For more details about the output files and reports, please refer to the +[output documentation](https://nf-co.re/rnadeseq/output). + +## Credits + +nf-core/rnadeseq was originally written by . + +We thank the following people for their extensive assistance in the development of this pipeline: + + + +## Contributions and Support + +If you would like to contribute to this pipeline, please see the [contributing guidelines](.github/CONTRIBUTING.md). + +For further information or help, don't hesitate to get in touch on the [Slack `#rnadeseq` channel](https://nfcore.slack.com/channels/rnadeseq) (you can join with [this invite](https://nf-co.re/join/slack)). + +## Citations + + + + + + +An extensive list of references for the tools used by the pipeline can be found in the [`CITATIONS.md`](CITATIONS.md) file. + +You can cite the `nf-core` publication as follows: + +> **The nf-core framework for community-curated bioinformatics pipelines.** +> +> Philip Ewels, Alexander Peltzer, Sven Fillinger, Harshil Patel, Johannes Alneberg, Andreas Wilm, Maxime Ulysse Garcia, Paolo Di Tommaso & Sven Nahnsen. +> +> _Nat Biotechnol._ 2020 Feb 13. doi: [10.1038/s41587-020-0439-x](https://dx.doi.org/10.1038/s41587-020-0439-x). diff --git a/assets/adaptivecard.json b/assets/adaptivecard.json new file mode 100644 index 00000000..99786880 --- /dev/null +++ b/assets/adaptivecard.json @@ -0,0 +1,67 @@ +{ + "type": "message", + "attachments": [ + { + "contentType": "application/vnd.microsoft.card.adaptive", + "contentUrl": null, + "content": { + "\$schema": "http://adaptivecards.io/schemas/adaptive-card.json", + "msteams": { + "width": "Full" + }, + "type": "AdaptiveCard", + "version": "1.2", + "body": [ + { + "type": "TextBlock", + "size": "Large", + "weight": "Bolder", + "color": "<% if (success) { %>Good<% } else { %>Attention<%} %>", + "text": "nf-core/rnadeseq v${version} - ${runName}", + "wrap": true + }, + { + "type": "TextBlock", + "spacing": "None", + "text": "Completed at ${dateComplete} (duration: ${duration})", + "isSubtle": true, + "wrap": true + }, + { + "type": "TextBlock", + "text": "<% if (success) { %>Pipeline completed successfully!<% } else { %>Pipeline completed with errors. The full error message was: ${errorReport}.<% } %>", + "wrap": true + }, + { + "type": "TextBlock", + "text": "The command used to launch the workflow was as follows:", + "wrap": true + }, + { + "type": "TextBlock", + "text": "${commandLine}", + "isSubtle": true, + "wrap": true + } + ], + "actions": [ + { + "type": "Action.ShowCard", + "title": "Pipeline Configuration", + "card": { + "type": "AdaptiveCard", + "\$schema": "http://adaptivecards.io/schemas/adaptive-card.json", + "body": [ + { + "type": "FactSet", + "facts": [<% out << summary.collect{ k,v -> "{\"title\": \"$k\", \"value\" : \"$v\"}"}.join(",\n") %> + ] + } + ] + } + } + ] + } + } + ] +} diff --git a/assets/email_template.html b/assets/email_template.html new file mode 100644 index 00000000..55bcca05 --- /dev/null +++ b/assets/email_template.html @@ -0,0 +1,53 @@ + + + + + + + + nf-core/rnadeseq Pipeline Report + + + + + + diff --git a/assets/email_template.txt b/assets/email_template.txt new file mode 100644 index 00000000..9a27eebe --- /dev/null +++ b/assets/email_template.txt @@ -0,0 +1,39 @@ +---------------------------------------------------- + ,--./,-. + ___ __ __ __ ___ /,-._.--~\\ + |\\ | |__ __ / ` / \\ |__) |__ } { + | \\| | \\__, \\__/ | \\ |___ \\`-._,-`-, + `._,._,' + nf-core/rnadeseq ${version} +---------------------------------------------------- +Run Name: $runName + +<% if (success){ + out << "## nf-core/rnadeseq execution completed successfully! ##" +} else { + out << """#################################################### +## nf-core/rnadeseq execution completed unsuccessfully! ## +#################################################### +The exit status of the task that caused the workflow execution to fail was: $exitStatus. +The full error message was: + +${errorReport} +""" +} %> + + +The workflow was completed at $dateComplete (duration: $duration) + +The command used to launch the workflow was as follows: + + $commandLine + + + +Pipeline Configuration: +----------------------- +<% out << summary.collect{ k,v -> " - $k: $v" }.join("\n") %> + +-- +nf-core/rnadeseq +https://github.com/nf-core/rnadeseq diff --git a/assets/methods_description_template.yml b/assets/methods_description_template.yml new file mode 100644 index 00000000..04b860b4 --- /dev/null +++ b/assets/methods_description_template.yml @@ -0,0 +1,29 @@ +id: "nf-core-rnadeseq-methods-description" +description: "Suggested text and references to use when describing pipeline usage within the methods section of a publication." +section_name: "nf-core/rnadeseq Methods Description" +section_href: "https://github.com/nf-core/rnadeseq" +plot_type: "html" +## TODO nf-core: Update the HTML below to your preferred methods description, e.g. add publication citation for this pipeline +## You inject any metadata in the Nextflow '${workflow}' object +data: | +

Methods

+

Data was processed using nf-core/rnadeseq v${workflow.manifest.version} ${doi_text} of the nf-core collection of workflows (Ewels et al., 2020), utilising reproducible software environments from the Bioconda (Grüning et al., 2018) and Biocontainers (da Veiga Leprevost et al., 2017) projects.

+

The pipeline was executed with Nextflow v${workflow.nextflow.version} (Di Tommaso et al., 2017) with the following command:

+ 
${workflow.commandLine}
+

${tool_citations}

+

References

+
    +
  • Di Tommaso, P., Chatzou, M., Floden, E. W., Barja, P. P., Palumbo, E., & Notredame, C. (2017). Nextflow enables reproducible computational workflows. Nature Biotechnology, 35(4), 316-319. doi: 10.1038/nbt.3820
    • +
  • Ewels, P. A., Peltzer, A., Fillinger, S., Patel, H., Alneberg, J., Wilm, A., Garcia, M. U., Di Tommaso, P., & Nahnsen, S. (2020). The nf-core framework for community-curated bioinformatics pipelines. Nature Biotechnology, 38(3), 276-278. doi: 10.1038/s41587-020-0439-x
    • +
  • Grüning, B., Dale, R., Sjödin, A., Chapman, B. A., Rowe, J., Tomkins-Tinch, C. H., Valieris, R., Köster, J., & Bioconda Team. (2018). Bioconda: sustainable and comprehensive software distribution for the life sciences. Nature Methods, 15(7), 475–476. doi: 10.1038/s41592-018-0046-7
    • +
  • da Veiga Leprevost, F., Grüning, B. A., Alves Aflitos, S., Röst, H. L., Uszkoreit, J., Barsnes, H., Vaudel, M., Moreno, P., Gatto, L., Weber, J., Bai, M., Jimenez, R. C., Sachsenberg, T., Pfeuffer, J., Vera Alvarez, R., Griss, J., Nesvizhskii, A. I., & Perez-Riverol, Y. (2017). BioContainers: an open-source and community-driven framework for software standardization. Bioinformatics (Oxford, England), 33(16), 2580–2582. doi: 10.1093/bioinformatics/btx192
    • + ${tool_bibliography} +
+
+
Notes:
+
    + ${nodoi_text} +
  • The command above does not include parameters contained in any configs or profiles that may have been used. Ensure the config file is also uploaded with your publication!
    • +
  • You should also cite all software used within this run. Check the "Software Versions" of this report to get version information.
    • +
+
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For information about how to interpret these results, please see the + documentation. +report_section_order: + "nf-core-rnadeseq-methods-description": + order: -1000 + software_versions: + order: -1001 + "nf-core-rnadeseq-summary": + order: -1002 + +export_plots: true + +disable_version_detection: true diff --git a/assets/nf-core-rnadeseq_logo_light.png b/assets/nf-core-rnadeseq_logo_light.png new file mode 100644 index 0000000000000000000000000000000000000000..8cc8118fe073394641158572eb3ba56e1f2a6703 GIT binary patch literal 78987 zcmeEui93|*|NktQ%2p{POG2_GTh>fbF$vKkWvL`7S+eglWvLVyq^!vx5wgoR)sd*| zWoJr4wy}@>_kPs*ocb4j*Y~ijuVhOY*KysvQ|q$y*J-S zk72oMpYe0q?VAbaq#*9EMu$IdcoXp^so>_-$xz2=@*S~&`he~jv*V1qmOg7gB~|ku z4iN>q9{kdyZ|YpwtFN7@xuSZ(Q#H%X?NoYKwa=aXj}v2~+N64J89ds7jyopzTE&lL zTX=gIP6(U()S#j$694J$cS-q-%;cBHV%V6wo@O1Lq5Km$NZ4lj<2c zwqe?icigw3j2(aYEkjNIUZPcw=4bC$%q+7b?p1P^!$01``3%CuY^87 z;+7(ucG8n%c1WY$)*4{X-}BJ>bo;<}^v>;*r&@E0IG1l!Tp!}0zq857k7xHop;0Kp 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+Output files + +- `fastqc/` + - `*_fastqc.html`: FastQC report containing quality metrics. + - `*_fastqc.zip`: Zip archive containing the FastQC report, tab-delimited data file and plot images. + +
+ +[FastQC](http://www.bioinformatics.babraham.ac.uk/projects/fastqc/) gives general quality metrics about your sequenced reads. It provides information about the quality score distribution across your reads, per base sequence content (%A/T/G/C), adapter contamination and overrepresented sequences. For further reading and documentation see the [FastQC help pages](http://www.bioinformatics.babraham.ac.uk/projects/fastqc/Help/). + +![MultiQC - FastQC sequence counts plot](images/mqc_fastqc_counts.png) + +![MultiQC - FastQC mean quality scores plot](images/mqc_fastqc_quality.png) + +![MultiQC - FastQC adapter content plot](images/mqc_fastqc_adapter.png) + +:::note +The FastQC plots displayed in the MultiQC report shows _untrimmed_ reads. They may contain adapter sequence and potentially regions with low quality. +::: + +### MultiQC + +
+Output files + +- `multiqc/` + - `multiqc_report.html`: a standalone HTML file that can be viewed in your web browser. + - `multiqc_data/`: directory containing parsed statistics from the different tools used in the pipeline. + - `multiqc_plots/`: directory containing static images from the report in various formats. + +
+ +[MultiQC](http://multiqc.info) is a visualization tool that generates a single HTML report summarising all samples in your project. Most of the pipeline QC results are visualised in the report and further statistics are available in the report data directory. + +Results generated by MultiQC collate pipeline QC from supported tools e.g. FastQC. The pipeline has special steps which also allow the software versions to be reported in the MultiQC output for future traceability. For more information about how to use MultiQC reports, see . + +### Pipeline information + +
+Output files + +- `pipeline_info/` + - Reports generated by Nextflow: `execution_report.html`, `execution_timeline.html`, `execution_trace.txt` and `pipeline_dag.dot`/`pipeline_dag.svg`. + - Reports generated by the pipeline: `pipeline_report.html`, `pipeline_report.txt` and `software_versions.yml`. The `pipeline_report*` files will only be present if the `--email` / `--email_on_fail` parameter's are used when running the pipeline. + - Reformatted samplesheet files used as input to the pipeline: `samplesheet.valid.csv`. + - Parameters used by the pipeline run: `params.json`. + +
+ +[Nextflow](https://www.nextflow.io/docs/latest/tracing.html) provides excellent functionality for generating various reports relevant to the running and execution of the pipeline. This will allow you to troubleshoot errors with the running of the pipeline, and also provide you with other information such as launch commands, run times and resource usage. diff --git a/docs/usage.md b/docs/usage.md new file mode 100644 index 00000000..18df612e --- /dev/null +++ b/docs/usage.md @@ -0,0 +1,226 @@ +# nf-core/rnadeseq: Usage + +## :warning: Please read this documentation on the nf-core website: [https://nf-co.re/rnadeseq/usage](https://nf-co.re/rnadeseq/usage) + +> _Documentation of pipeline parameters is generated automatically from the pipeline schema and can no longer be found in markdown files._ + +## Introduction + + + +## Samplesheet input + +You will need to create a samplesheet with information about the samples you would like to analyse before running the pipeline. Use this parameter to specify its location. It has to be a comma-separated file with 3 columns, and a header row as shown in the examples below. + +```bash +--input '[path to samplesheet file]' +``` + +### Multiple runs of the same sample + +The `sample` identifiers have to be the same when you have re-sequenced the same sample more than once e.g. to increase sequencing depth. The pipeline will concatenate the raw reads before performing any downstream analysis. Below is an example for the same sample sequenced across 3 lanes: + +```csv title="samplesheet.csv" +sample,fastq_1,fastq_2 +CONTROL_REP1,AEG588A1_S1_L002_R1_001.fastq.gz,AEG588A1_S1_L002_R2_001.fastq.gz +CONTROL_REP1,AEG588A1_S1_L003_R1_001.fastq.gz,AEG588A1_S1_L003_R2_001.fastq.gz +CONTROL_REP1,AEG588A1_S1_L004_R1_001.fastq.gz,AEG588A1_S1_L004_R2_001.fastq.gz +``` + +### Full samplesheet + +The pipeline will auto-detect whether a sample is single- or paired-end using the information provided in the samplesheet. The samplesheet can have as many columns as you desire, however, there is a strict requirement for the first 3 columns to match those defined in the table below. + +A final samplesheet file consisting of both single- and paired-end data may look something like the one below. This is for 6 samples, where `TREATMENT_REP3` has been sequenced twice. + +```csv title="samplesheet.csv" +sample,fastq_1,fastq_2 +CONTROL_REP1,AEG588A1_S1_L002_R1_001.fastq.gz,AEG588A1_S1_L002_R2_001.fastq.gz +CONTROL_REP2,AEG588A2_S2_L002_R1_001.fastq.gz,AEG588A2_S2_L002_R2_001.fastq.gz +CONTROL_REP3,AEG588A3_S3_L002_R1_001.fastq.gz,AEG588A3_S3_L002_R2_001.fastq.gz +TREATMENT_REP1,AEG588A4_S4_L003_R1_001.fastq.gz, +TREATMENT_REP2,AEG588A5_S5_L003_R1_001.fastq.gz, +TREATMENT_REP3,AEG588A6_S6_L003_R1_001.fastq.gz, +TREATMENT_REP3,AEG588A6_S6_L004_R1_001.fastq.gz, +``` + +| Column | Description | +| --------- | -------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | +| `sample` | Custom sample name. This entry will be identical for multiple sequencing libraries/runs from the same sample. Spaces in sample names are automatically converted to underscores (`_`). | +| `fastq_1` | Full path to FastQ file for Illumina short reads 1. File has to be gzipped and have the extension ".fastq.gz" or ".fq.gz". | +| `fastq_2` | Full path to FastQ file for Illumina short reads 2. File has to be gzipped and have the extension ".fastq.gz" or ".fq.gz". | + +An [example samplesheet](../assets/samplesheet.csv) has been provided with the pipeline. + +## Running the pipeline + +The typical command for running the pipeline is as follows: + +```bash +nextflow run nf-core/rnadeseq --input ./samplesheet.csv --outdir ./results --genome GRCh37 -profile docker +``` + +This will launch the pipeline with the `docker` configuration profile. See below for more information about profiles. + +Note that the pipeline will create the following files in your working directory: + +```bash +work # Directory containing the nextflow working files + # Finished results in specified location (defined with --outdir) +.nextflow_log # Log file from Nextflow +# Other nextflow hidden files, eg. history of pipeline runs and old logs. +``` + +If you wish to repeatedly use the same parameters for multiple runs, rather than specifying each flag in the command, you can specify these in a params file. + +Pipeline settings can be provided in a `yaml` or `json` file via `-params-file `. + +:::warning +Do not use `-c ` to specify parameters as this will result in errors. Custom config files specified with `-c` must only be used for [tuning process resource specifications](https://nf-co.re/docs/usage/configuration#tuning-workflow-resources), other infrastructural tweaks (such as output directories), or module arguments (args). +::: + +The above pipeline run specified with a params file in yaml format: + +```bash +nextflow run nf-core/rnadeseq -profile docker -params-file params.yaml +``` + +with `params.yaml` containing: + +```yaml +input: './samplesheet.csv' +outdir: './results/' +genome: 'GRCh37' +<...> +``` + +You can also generate such `YAML`/`JSON` files via [nf-core/launch](https://nf-co.re/launch). + +### Updating the pipeline + +When you run the above command, Nextflow automatically pulls the pipeline code from GitHub and stores it as a cached version. When running the pipeline after this, it will always use the cached version if available - even if the pipeline has been updated since. To make sure that you're running the latest version of the pipeline, make sure that you regularly update the cached version of the pipeline: + +```bash +nextflow pull nf-core/rnadeseq +``` + +### Reproducibility + +It is a good idea to specify a pipeline version when running the pipeline on your data. This ensures that a specific version of the pipeline code and software are used when you run your pipeline. If you keep using the same tag, you'll be running the same version of the pipeline, even if there have been changes to the code since. + +First, go to the [nf-core/rnadeseq releases page](https://github.com/nf-core/rnadeseq/releases) and find the latest pipeline version - numeric only (eg. `1.3.1`). Then specify this when running the pipeline with `-r` (one hyphen) - eg. `-r 1.3.1`. Of course, you can switch to another version by changing the number after the `-r` flag. + +This version number will be logged in reports when you run the pipeline, so that you'll know what you used when you look back in the future. For example, at the bottom of the MultiQC reports. + +To further assist in reproducbility, you can use share and re-use [parameter files](#running-the-pipeline) to repeat pipeline runs with the same settings without having to write out a command with every single parameter. + +:::tip +If you wish to share such profile (such as upload as supplementary material for academic publications), make sure to NOT include cluster specific paths to files, nor institutional specific profiles. +::: + +## Core Nextflow arguments + +:::note +These options are part of Nextflow and use a _single_ hyphen (pipeline parameters use a double-hyphen). +::: + +### `-profile` + +Use this parameter to choose a configuration profile. Profiles can give configuration presets for different compute environments. + +Several generic profiles are bundled with the pipeline which instruct the pipeline to use software packaged using different methods (Docker, Singularity, Podman, Shifter, Charliecloud, Apptainer, Conda) - see below. + +:::info +We highly recommend the use of Docker or Singularity containers for full pipeline reproducibility, however when this is not possible, Conda is also supported. +::: + +The pipeline also dynamically loads configurations from [https://github.com/nf-core/configs](https://github.com/nf-core/configs) when it runs, making multiple config profiles for various institutional clusters available at run time. For more information and to see if your system is available in these configs please see the [nf-core/configs documentation](https://github.com/nf-core/configs#documentation). + +Note that multiple profiles can be loaded, for example: `-profile test,docker` - the order of arguments is important! +They are loaded in sequence, so later profiles can overwrite earlier profiles. + +If `-profile` is not specified, the pipeline will run locally and expect all software to be installed and available on the `PATH`. This is _not_ recommended, since it can lead to different results on different machines dependent on the computer enviroment. + +- `test` + - A profile with a complete configuration for automated testing + - Includes links to test data so needs no other parameters +- `docker` + - A generic configuration profile to be used with [Docker](https://docker.com/) +- `singularity` + - A generic configuration profile to be used with [Singularity](https://sylabs.io/docs/) +- `podman` + - A generic configuration profile to be used with [Podman](https://podman.io/) +- `shifter` + - A generic configuration profile to be used with [Shifter](https://nersc.gitlab.io/development/shifter/how-to-use/) +- `charliecloud` + - A generic configuration profile to be used with [Charliecloud](https://hpc.github.io/charliecloud/) +- `apptainer` + - A generic configuration profile to be used with [Apptainer](https://apptainer.org/) +- `wave` + - A generic configuration profile to enable [Wave](https://seqera.io/wave/) containers. Use together with one of the above (requires Nextflow ` 24.03.0-edge` or later). +- `conda` + - A generic configuration profile to be used with [Conda](https://conda.io/docs/). Please only use Conda as a last resort i.e. when it's not possible to run the pipeline with Docker, Singularity, Podman, Shifter, Charliecloud, or Apptainer. + +### `-resume` + +Specify this when restarting a pipeline. Nextflow will use cached results from any pipeline steps where the inputs are the same, continuing from where it got to previously. For input to be considered the same, not only the names must be identical but the files' contents as well. For more info about this parameter, see [this blog post](https://www.nextflow.io/blog/2019/demystifying-nextflow-resume.html). + +You can also supply a run name to resume a specific run: `-resume [run-name]`. Use the `nextflow log` command to show previous run names. + +### `-c` + +Specify the path to a specific config file (this is a core Nextflow command). See the [nf-core website documentation](https://nf-co.re/usage/configuration) for more information. + +## Custom configuration + +### Resource requests + +Whilst the default requirements set within the pipeline will hopefully work for most people and with most input data, you may find that you want to customise the compute resources that the pipeline requests. Each step in the pipeline has a default set of requirements for number of CPUs, memory and time. For most of the steps in the pipeline, if the job exits with any of the error codes specified [here](https://github.com/nf-core/rnaseq/blob/4c27ef5610c87db00c3c5a3eed10b1d161abf575/conf/base.config#L18) it will automatically be resubmitted with higher requests (2 x original, then 3 x original). If it still fails after the third attempt then the pipeline execution is stopped. + +To change the resource requests, please see the [max resources](https://nf-co.re/docs/usage/configuration#max-resources) and [tuning workflow resources](https://nf-co.re/docs/usage/configuration#tuning-workflow-resources) section of the nf-core website. + +### Custom Containers + +In some cases you may wish to change which container or conda environment a step of the pipeline uses for a particular tool. By default nf-core pipelines use containers and software from the [biocontainers](https://biocontainers.pro/) or [bioconda](https://bioconda.github.io/) projects. However in some cases the pipeline specified version maybe out of date. + +To use a different container from the default container or conda environment specified in a pipeline, please see the [updating tool versions](https://nf-co.re/docs/usage/configuration#updating-tool-versions) section of the nf-core website. + +### Custom Tool Arguments + +A pipeline might not always support every possible argument or option of a particular tool used in pipeline. Fortunately, nf-core pipelines provide some freedom to users to insert additional parameters that the pipeline does not include by default. + +To learn how to provide additional arguments to a particular tool of the pipeline, please see the [customising tool arguments](https://nf-co.re/docs/usage/configuration#customising-tool-arguments) section of the nf-core website. + +### nf-core/configs + +In most cases, you will only need to create a custom config as a one-off but if you and others within your organisation are likely to be running nf-core pipelines regularly and need to use the same settings regularly it may be a good idea to request that your custom config file is uploaded to the `nf-core/configs` git repository. Before you do this please can you test that the config file works with your pipeline of choice using the `-c` parameter. You can then create a pull request to the `nf-core/configs` repository with the addition of your config file, associated documentation file (see examples in [`nf-core/configs/docs`](https://github.com/nf-core/configs/tree/master/docs)), and amending [`nfcore_custom.config`](https://github.com/nf-core/configs/blob/master/nfcore_custom.config) to include your custom profile. + +See the main [Nextflow documentation](https://www.nextflow.io/docs/latest/config.html) for more information about creating your own configuration files. + +If you have any questions or issues please send us a message on [Slack](https://nf-co.re/join/slack) on the [`#configs` channel](https://nfcore.slack.com/channels/configs). + +## Azure Resource Requests + +To be used with the `azurebatch` profile by specifying the `-profile azurebatch`. +We recommend providing a compute `params.vm_type` of `Standard_D16_v3` VMs by default but these options can be changed if required. + +Note that the choice of VM size depends on your quota and the overall workload during the analysis. +For a thorough list, please refer the [Azure Sizes for virtual machines in Azure](https://docs.microsoft.com/en-us/azure/virtual-machines/sizes). + +## Running in the background + +Nextflow handles job submissions and supervises the running jobs. The Nextflow process must run until the pipeline is finished. + +The Nextflow `-bg` flag launches Nextflow in the background, detached from your terminal so that the workflow does not stop if you log out of your session. The logs are saved to a file. + +Alternatively, you can use `screen` / `tmux` or similar tool to create a detached session which you can log back into at a later time. +Some HPC setups also allow you to run nextflow within a cluster job submitted your job scheduler (from where it submits more jobs). + +## Nextflow memory requirements + +In some cases, the Nextflow Java virtual machines can start to request a large amount of memory. +We recommend adding the following line to your environment to limit this (typically in `~/.bashrc` or `~./bash_profile`): + +```bash +NXF_OPTS='-Xms1g -Xmx4g' +``` diff --git a/main.nf b/main.nf new file mode 100644 index 00000000..efd9546b --- /dev/null +++ b/main.nf @@ -0,0 +1,112 @@ +#!/usr/bin/env nextflow +/* +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ + nf-core/rnadeseq +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ + Github : https://github.com/nf-core/rnadeseq + Website: https://nf-co.re/rnadeseq + Slack : https://nfcore.slack.com/channels/rnadeseq +---------------------------------------------------------------------------------------- +*/ + +nextflow.enable.dsl = 2 + +/* +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ + IMPORT FUNCTIONS / MODULES / SUBWORKFLOWS / WORKFLOWS +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ +*/ + +include { RNADESEQ } from './workflows/rnadeseq' +include { PIPELINE_INITIALISATION } from './subworkflows/local/utils_nfcore_rnadeseq_pipeline' +include { PIPELINE_COMPLETION } from './subworkflows/local/utils_nfcore_rnadeseq_pipeline' + +include { getGenomeAttribute } from './subworkflows/local/utils_nfcore_rnadeseq_pipeline' + +/* +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ + GENOME PARAMETER VALUES +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ +*/ + +// TODO nf-core: Remove this line if you don't need a FASTA file +// This is an example of how to use getGenomeAttribute() to fetch parameters +// from igenomes.config using `--genome` +params.fasta = getGenomeAttribute('fasta') + +/* +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ + NAMED WORKFLOWS FOR PIPELINE +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ +*/ + +// +// WORKFLOW: Run main analysis pipeline depending on type of input +// +workflow NFCORE_RNADESEQ { + + take: + samplesheet // channel: samplesheet read in from --input + + main: + + // + // WORKFLOW: Run pipeline + // + RNADESEQ ( + samplesheet + ) + + emit: + multiqc_report = RNADESEQ.out.multiqc_report // channel: /path/to/multiqc_report.html + +} +/* +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ + RUN MAIN WORKFLOW +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ +*/ + +workflow { + + main: + + // + // SUBWORKFLOW: Run initialisation tasks + // + PIPELINE_INITIALISATION ( + params.version, + params.help, + params.validate_params, + params.monochrome_logs, + args, + params.outdir, + params.input + ) + + // + // WORKFLOW: Run main workflow + // + NFCORE_RNADESEQ ( + PIPELINE_INITIALISATION.out.samplesheet + ) + + // + // SUBWORKFLOW: Run completion tasks + // + PIPELINE_COMPLETION ( + params.email, + params.email_on_fail, + params.plaintext_email, + params.outdir, + params.monochrome_logs, + params.hook_url, + NFCORE_RNADESEQ.out.multiqc_report + ) +} + +/* +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ + THE END +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ +*/ diff --git a/modules.json b/modules.json new file mode 100644 index 00000000..1e166e57 --- /dev/null +++ b/modules.json @@ -0,0 +1,41 @@ +{ + "name": "nf-core/rnadeseq", + "homePage": "https://github.com/nf-core/rnadeseq", + "repos": { + "https://github.com/nf-core/modules.git": { + "modules": { + "nf-core": { + "fastqc": { + "branch": "master", + "git_sha": "285a50500f9e02578d90b3ce6382ea3c30216acd", + "installed_by": ["modules"] + }, + "multiqc": { + "branch": "master", + "git_sha": "b7ebe95761cd389603f9cc0e0dc384c0f663815a", + "installed_by": ["modules"] + } + } + }, + "subworkflows": { + "nf-core": { + "utils_nextflow_pipeline": { + "branch": "master", + "git_sha": "5caf7640a9ef1d18d765d55339be751bb0969dfa", + "installed_by": ["subworkflows"] + }, + "utils_nfcore_pipeline": { + "branch": "master", + "git_sha": "92de218a329bfc9a9033116eb5f65fd270e72ba3", + "installed_by": ["subworkflows"] + }, + "utils_nfvalidation_plugin": { + "branch": "master", + "git_sha": "5caf7640a9ef1d18d765d55339be751bb0969dfa", + "installed_by": ["subworkflows"] + } + } + } + } + } +} diff --git a/modules/nf-core/fastqc/environment.yml b/modules/nf-core/fastqc/environment.yml new file mode 100644 index 00000000..1787b38a --- /dev/null +++ b/modules/nf-core/fastqc/environment.yml @@ -0,0 +1,7 @@ +name: fastqc +channels: + - conda-forge + - bioconda + - defaults +dependencies: + - bioconda::fastqc=0.12.1 diff --git a/modules/nf-core/fastqc/main.nf b/modules/nf-core/fastqc/main.nf new file mode 100644 index 00000000..d79f1c86 --- /dev/null +++ b/modules/nf-core/fastqc/main.nf @@ -0,0 +1,61 @@ +process FASTQC { + tag "$meta.id" + label 'process_medium' + + conda "${moduleDir}/environment.yml" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/fastqc:0.12.1--hdfd78af_0' : + 'biocontainers/fastqc:0.12.1--hdfd78af_0' }" + + input: + tuple val(meta), path(reads) + + output: + tuple val(meta), path("*.html"), emit: html + tuple val(meta), path("*.zip") , emit: zip + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + // Make list of old name and new name pairs to use for renaming in the bash while loop + def old_new_pairs = reads instanceof Path || reads.size() == 1 ? [[ reads, "${prefix}.${reads.extension}" ]] : reads.withIndex().collect { entry, index -> [ entry, "${prefix}_${index + 1}.${entry.extension}" ] } + def rename_to = old_new_pairs*.join(' ').join(' ') + def renamed_files = old_new_pairs.collect{ old_name, new_name -> new_name }.join(' ') + + def memory_in_mb = MemoryUnit.of("${task.memory}").toUnit('MB') + // FastQC memory value allowed range (100 - 10000) + def fastqc_memory = memory_in_mb > 10000 ? 10000 : (memory_in_mb < 100 ? 100 : memory_in_mb) + + """ + printf "%s %s\\n" $rename_to | while read old_name new_name; do + [ -f "\${new_name}" ] || ln -s \$old_name \$new_name + done + + fastqc \\ + $args \\ + --threads $task.cpus \\ + --memory $fastqc_memory \\ + $renamed_files + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + fastqc: \$( fastqc --version | sed '/FastQC v/!d; s/.*v//' ) + END_VERSIONS + """ + + stub: + def prefix = task.ext.prefix ?: "${meta.id}" + """ + touch ${prefix}.html + touch ${prefix}.zip + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + fastqc: \$( fastqc --version | sed '/FastQC v/!d; s/.*v//' ) + END_VERSIONS + """ +} diff --git a/modules/nf-core/fastqc/meta.yml b/modules/nf-core/fastqc/meta.yml new file mode 100644 index 00000000..ee5507e0 --- /dev/null +++ b/modules/nf-core/fastqc/meta.yml @@ -0,0 +1,57 @@ +name: fastqc +description: Run FastQC on sequenced reads +keywords: + - quality control + - qc + - adapters + - fastq +tools: + - fastqc: + description: | + FastQC gives general quality metrics about your reads. + It provides information about the quality score distribution + across your reads, the per base sequence content (%A/C/G/T). + You get information about adapter contamination and other + overrepresented sequences. + homepage: https://www.bioinformatics.babraham.ac.uk/projects/fastqc/ + documentation: https://www.bioinformatics.babraham.ac.uk/projects/fastqc/Help/ + licence: ["GPL-2.0-only"] +input: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - reads: + type: file + description: | + List of input FastQ files of size 1 and 2 for single-end and paired-end data, + respectively. +output: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - html: + type: file + description: FastQC report + pattern: "*_{fastqc.html}" + - zip: + type: file + description: FastQC report archive + pattern: "*_{fastqc.zip}" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" +authors: + - "@drpatelh" + - "@grst" + - "@ewels" + - "@FelixKrueger" +maintainers: + - "@drpatelh" + - "@grst" + - "@ewels" + - "@FelixKrueger" diff --git a/modules/nf-core/fastqc/tests/main.nf.test b/modules/nf-core/fastqc/tests/main.nf.test new file mode 100644 index 00000000..70edae4d --- /dev/null +++ b/modules/nf-core/fastqc/tests/main.nf.test @@ -0,0 +1,212 @@ +nextflow_process { + + name "Test Process FASTQC" + script "../main.nf" + process "FASTQC" + + tag "modules" + tag "modules_nfcore" + tag "fastqc" + + test("sarscov2 single-end [fastq]") { + + when { + process { + """ + input[0] = Channel.of([ + [ id: 'test', single_end:true ], + [ file(params.modules_testdata_base_path + 'genomics/sarscov2/illumina/fastq/test_1.fastq.gz', checkIfExists: true) ] + ]) + """ + } + } + + then { + assertAll ( + { assert process.success }, + + // NOTE The report contains the date inside it, which means that the md5sum is stable per day, but not longer than that. So you can't md5sum it. + // looks like this:
Mon 2 Oct 2023
test.gz
+ // https://github.com/nf-core/modules/pull/3903#issuecomment-1743620039 + + { assert process.out.html[0][1] ==~ ".*/test_fastqc.html" }, + { assert process.out.zip[0][1] ==~ ".*/test_fastqc.zip" }, + { assert path(process.out.html[0][1]).text.contains("File typeConventional base calls") }, + + { assert snapshot(process.out.versions).match("fastqc_versions_single") } + ) + } + } + + test("sarscov2 paired-end [fastq]") { + + when { + process { + """ + input[0] = Channel.of([ + [id: 'test', single_end: false], // meta map + [ file(params.modules_testdata_base_path + 'genomics/sarscov2/illumina/fastq/test_1.fastq.gz', checkIfExists: true), + file(params.modules_testdata_base_path + 'genomics/sarscov2/illumina/fastq/test_2.fastq.gz', checkIfExists: true) ] + ]) + """ + } + } + + then { + assertAll ( + { assert process.success }, + + { assert process.out.html[0][1][0] ==~ ".*/test_1_fastqc.html" }, + { assert process.out.html[0][1][1] ==~ ".*/test_2_fastqc.html" }, + { assert process.out.zip[0][1][0] ==~ ".*/test_1_fastqc.zip" }, + { assert process.out.zip[0][1][1] ==~ ".*/test_2_fastqc.zip" }, + { assert path(process.out.html[0][1][0]).text.contains("File typeConventional base calls") }, + { assert path(process.out.html[0][1][1]).text.contains("File typeConventional base calls") }, + + { assert snapshot(process.out.versions).match("fastqc_versions_paired") } + ) + } + } + + test("sarscov2 interleaved [fastq]") { + + when { + process { + """ + input[0] = Channel.of([ + [id: 'test', single_end: false], // meta map + file(params.modules_testdata_base_path + 'genomics/sarscov2/illumina/fastq/test_interleaved.fastq.gz', checkIfExists: true) + ]) + """ + } + } + + then { + assertAll ( + { assert process.success }, + + { assert process.out.html[0][1] ==~ ".*/test_fastqc.html" }, + { assert process.out.zip[0][1] ==~ ".*/test_fastqc.zip" }, + { assert path(process.out.html[0][1]).text.contains("File typeConventional base calls") }, + + { assert snapshot(process.out.versions).match("fastqc_versions_interleaved") } + ) + } + } + + test("sarscov2 paired-end [bam]") { + + when { + process { + """ + input[0] = Channel.of([ + [id: 'test', single_end: false], // meta map + file(params.modules_testdata_base_path + 'genomics/sarscov2/illumina/bam/test.paired_end.sorted.bam', checkIfExists: true) + ]) + """ + } + } + + then { + assertAll ( + { assert process.success }, + + { assert process.out.html[0][1] ==~ ".*/test_fastqc.html" }, + { assert process.out.zip[0][1] ==~ ".*/test_fastqc.zip" }, + { assert path(process.out.html[0][1]).text.contains("File typeConventional base calls") }, + + { assert snapshot(process.out.versions).match("fastqc_versions_bam") } + ) + } + } + + test("sarscov2 multiple [fastq]") { + + when { + process { + """ + input[0] = Channel.of([ + [id: 'test', single_end: false], // meta map + [ file(params.modules_testdata_base_path + 'genomics/sarscov2/illumina/fastq/test_1.fastq.gz', checkIfExists: true), + file(params.modules_testdata_base_path + 'genomics/sarscov2/illumina/fastq/test_2.fastq.gz', checkIfExists: true), + file(params.modules_testdata_base_path + 'genomics/sarscov2/illumina/fastq/test2_1.fastq.gz', checkIfExists: true), + file(params.modules_testdata_base_path + 'genomics/sarscov2/illumina/fastq/test2_2.fastq.gz', checkIfExists: true) ] + ]) + """ + } + } + + then { + assertAll ( + { assert process.success }, + + { assert process.out.html[0][1][0] ==~ ".*/test_1_fastqc.html" }, + { assert process.out.html[0][1][1] ==~ ".*/test_2_fastqc.html" }, + { assert process.out.html[0][1][2] ==~ ".*/test_3_fastqc.html" }, + { assert process.out.html[0][1][3] ==~ ".*/test_4_fastqc.html" }, + { assert process.out.zip[0][1][0] ==~ ".*/test_1_fastqc.zip" }, + { assert process.out.zip[0][1][1] ==~ ".*/test_2_fastqc.zip" }, + { assert process.out.zip[0][1][2] ==~ ".*/test_3_fastqc.zip" }, + { assert process.out.zip[0][1][3] ==~ ".*/test_4_fastqc.zip" }, + { assert path(process.out.html[0][1][0]).text.contains("File typeConventional base calls") }, + { assert path(process.out.html[0][1][1]).text.contains("File typeConventional base calls") }, + { assert path(process.out.html[0][1][2]).text.contains("File typeConventional base calls") }, + { assert path(process.out.html[0][1][3]).text.contains("File typeConventional base calls") }, + + { assert snapshot(process.out.versions).match("fastqc_versions_multiple") } + ) + } + } + + test("sarscov2 custom_prefix") { + + when { + process { + """ + input[0] = Channel.of([ + [ id:'mysample', single_end:true ], // meta map + file(params.modules_testdata_base_path + 'genomics/sarscov2/illumina/fastq/test_1.fastq.gz', checkIfExists: true) + ]) + """ + } + } + + then { + assertAll ( + { assert process.success }, + + { assert process.out.html[0][1] ==~ ".*/mysample_fastqc.html" }, + { assert process.out.zip[0][1] ==~ ".*/mysample_fastqc.zip" }, + { assert path(process.out.html[0][1]).text.contains("File typeConventional base calls") }, + + { assert snapshot(process.out.versions).match("fastqc_versions_custom_prefix") } + ) + } + } + + test("sarscov2 single-end [fastq] - stub") { + + options "-stub" + + when { + process { + """ + input[0] = Channel.of([ + [ id: 'test', single_end:true ], + [ file(params.modules_testdata_base_path + 'genomics/sarscov2/illumina/fastq/test_1.fastq.gz', checkIfExists: true) ] + ]) + """ + } + } + + then { + assertAll ( + { assert process.success }, + { assert snapshot(process.out.html.collect { file(it[1]).getName() } + + process.out.zip.collect { file(it[1]).getName() } + + process.out.versions ).match("fastqc_stub") } + ) + } + } + +} diff --git a/modules/nf-core/fastqc/tests/main.nf.test.snap b/modules/nf-core/fastqc/tests/main.nf.test.snap new file mode 100644 index 00000000..86f7c311 --- /dev/null +++ b/modules/nf-core/fastqc/tests/main.nf.test.snap @@ -0,0 +1,88 @@ +{ + "fastqc_versions_interleaved": { + "content": [ + [ + "versions.yml:md5,e1cc25ca8af856014824abd842e93978" + ] + ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.10.1" + }, + "timestamp": "2024-01-31T17:40:07.293713" + }, + "fastqc_stub": { + "content": [ + [ + "test.html", + "test.zip", + "versions.yml:md5,e1cc25ca8af856014824abd842e93978" + ] + ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.10.1" + }, + "timestamp": "2024-01-31T17:31:01.425198" + }, + "fastqc_versions_multiple": { + "content": [ + [ + "versions.yml:md5,e1cc25ca8af856014824abd842e93978" + ] + ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.10.1" + }, + "timestamp": "2024-01-31T17:40:55.797907" + }, + "fastqc_versions_bam": { + "content": [ + [ + "versions.yml:md5,e1cc25ca8af856014824abd842e93978" + ] + ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.10.1" + }, + "timestamp": "2024-01-31T17:40:26.795862" + }, + "fastqc_versions_single": { + "content": [ + [ + "versions.yml:md5,e1cc25ca8af856014824abd842e93978" + ] + ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.10.1" + }, + "timestamp": "2024-01-31T17:39:27.043675" + }, + "fastqc_versions_paired": { + "content": [ + [ + "versions.yml:md5,e1cc25ca8af856014824abd842e93978" + ] + ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.10.1" + }, + "timestamp": "2024-01-31T17:39:47.584191" + }, + "fastqc_versions_custom_prefix": { + "content": [ + [ + "versions.yml:md5,e1cc25ca8af856014824abd842e93978" + ] + ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.10.1" + }, + "timestamp": "2024-01-31T17:41:14.576531" + } +} \ No newline at end of file diff --git a/modules/nf-core/fastqc/tests/tags.yml b/modules/nf-core/fastqc/tests/tags.yml new file mode 100644 index 00000000..7834294b --- /dev/null +++ b/modules/nf-core/fastqc/tests/tags.yml @@ -0,0 +1,2 @@ +fastqc: + - modules/nf-core/fastqc/** diff --git a/modules/nf-core/multiqc/environment.yml b/modules/nf-core/multiqc/environment.yml new file mode 100644 index 00000000..ca39fb67 --- /dev/null +++ b/modules/nf-core/multiqc/environment.yml @@ -0,0 +1,7 @@ +name: multiqc +channels: + - conda-forge + - bioconda + - defaults +dependencies: + - bioconda::multiqc=1.21 diff --git a/modules/nf-core/multiqc/main.nf b/modules/nf-core/multiqc/main.nf new file mode 100644 index 00000000..47ac352f --- /dev/null +++ b/modules/nf-core/multiqc/main.nf @@ -0,0 +1,55 @@ +process MULTIQC { + label 'process_single' + + conda "${moduleDir}/environment.yml" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/multiqc:1.21--pyhdfd78af_0' : + 'biocontainers/multiqc:1.21--pyhdfd78af_0' }" + + input: + path multiqc_files, stageAs: "?/*" + path(multiqc_config) + path(extra_multiqc_config) + path(multiqc_logo) + + output: + path "*multiqc_report.html", emit: report + path "*_data" , emit: data + path "*_plots" , optional:true, emit: plots + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def config = multiqc_config ? "--config $multiqc_config" : '' + def extra_config = extra_multiqc_config ? "--config $extra_multiqc_config" : '' + def logo = multiqc_logo ? /--cl-config 'custom_logo: "${multiqc_logo}"'/ : '' + """ + multiqc \\ + --force \\ + $args \\ + $config \\ + $extra_config \\ + $logo \\ + . + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + multiqc: \$( multiqc --version | sed -e "s/multiqc, version //g" ) + END_VERSIONS + """ + + stub: + """ + mkdir multiqc_data + touch multiqc_plots + touch multiqc_report.html + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + multiqc: \$( multiqc --version | sed -e "s/multiqc, version //g" ) + END_VERSIONS + """ +} diff --git a/modules/nf-core/multiqc/meta.yml b/modules/nf-core/multiqc/meta.yml new file mode 100644 index 00000000..45a9bc35 --- /dev/null +++ b/modules/nf-core/multiqc/meta.yml @@ -0,0 +1,58 @@ +name: multiqc +description: Aggregate results from bioinformatics analyses across many samples into a single report +keywords: + - QC + - bioinformatics tools + - Beautiful stand-alone HTML report +tools: + - multiqc: + description: | + MultiQC searches a given directory for analysis logs and compiles a HTML report. + It's a general use tool, perfect for summarising the output from numerous bioinformatics tools. + homepage: https://multiqc.info/ + documentation: https://multiqc.info/docs/ + licence: ["GPL-3.0-or-later"] +input: + - multiqc_files: + type: file + description: | + List of reports / files recognised by MultiQC, for example the html and zip output of FastQC + - multiqc_config: + type: file + description: Optional config yml for MultiQC + pattern: "*.{yml,yaml}" + - extra_multiqc_config: + type: file + description: Second optional config yml for MultiQC. Will override common sections in multiqc_config. + pattern: "*.{yml,yaml}" + - multiqc_logo: + type: file + description: Optional logo file for MultiQC + pattern: "*.{png}" +output: + - report: + type: file + description: MultiQC report file + pattern: "multiqc_report.html" + - data: + type: directory + description: MultiQC data dir + pattern: "multiqc_data" + - plots: + type: file + description: Plots created by MultiQC + pattern: "*_data" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" +authors: + - "@abhi18av" + - "@bunop" + - "@drpatelh" + - "@jfy133" +maintainers: + - "@abhi18av" + - "@bunop" + - "@drpatelh" + - "@jfy133" diff --git a/modules/nf-core/multiqc/tests/main.nf.test b/modules/nf-core/multiqc/tests/main.nf.test new file mode 100644 index 00000000..f1c4242e --- /dev/null +++ b/modules/nf-core/multiqc/tests/main.nf.test @@ -0,0 +1,84 @@ +nextflow_process { + + name "Test Process MULTIQC" + script "../main.nf" + process "MULTIQC" + + tag "modules" + tag "modules_nfcore" + tag "multiqc" + + test("sarscov2 single-end [fastqc]") { + + when { + process { + """ + input[0] = Channel.of(file(params.modules_testdata_base_path + 'genomics/sarscov2/illumina/fastqc/test_fastqc.zip', checkIfExists: true)) + input[1] = [] + input[2] = [] + input[3] = [] + """ + } + } + + then { + assertAll( + { assert process.success }, + { assert process.out.report[0] ==~ ".*/multiqc_report.html" }, + { assert process.out.data[0] ==~ ".*/multiqc_data" }, + { assert snapshot(process.out.versions).match("multiqc_versions_single") } + ) + } + + } + + test("sarscov2 single-end [fastqc] [config]") { + + when { + process { + """ + input[0] = Channel.of(file(params.modules_testdata_base_path + 'genomics/sarscov2/illumina/fastqc/test_fastqc.zip', checkIfExists: true)) + input[1] = Channel.of(file("https://github.com/nf-core/tools/raw/dev/nf_core/pipeline-template/assets/multiqc_config.yml", checkIfExists: true)) + input[2] = [] + input[3] = [] + """ + } + } + + then { + assertAll( + { assert process.success }, + { assert process.out.report[0] ==~ ".*/multiqc_report.html" }, + { assert process.out.data[0] ==~ ".*/multiqc_data" }, + { assert snapshot(process.out.versions).match("multiqc_versions_config") } + ) + } + } + + test("sarscov2 single-end [fastqc] - stub") { + + options "-stub" + + when { + process { + """ + input[0] = Channel.of(file(params.modules_testdata_base_path + 'genomics/sarscov2/illumina/fastqc/test_fastqc.zip', checkIfExists: true)) + input[1] = [] + input[2] = [] + input[3] = [] + """ + } + } + + then { + assertAll( + { assert process.success }, + { assert snapshot(process.out.report.collect { file(it).getName() } + + process.out.data.collect { file(it).getName() } + + process.out.plots.collect { file(it).getName() } + + process.out.versions ).match("multiqc_stub") } + ) + } + + } +} diff --git a/modules/nf-core/multiqc/tests/main.nf.test.snap b/modules/nf-core/multiqc/tests/main.nf.test.snap new file mode 100644 index 00000000..bfebd802 --- /dev/null +++ b/modules/nf-core/multiqc/tests/main.nf.test.snap @@ -0,0 +1,41 @@ +{ + "multiqc_versions_single": { + "content": [ + [ + "versions.yml:md5,21f35ee29416b9b3073c28733efe4b7d" + ] + ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.10.1" + }, + "timestamp": "2024-02-29T08:48:55.657331" + }, + "multiqc_stub": { + "content": [ + [ + "multiqc_report.html", + "multiqc_data", + "multiqc_plots", + "versions.yml:md5,21f35ee29416b9b3073c28733efe4b7d" + ] + ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.10.1" + }, + "timestamp": "2024-02-29T08:49:49.071937" + }, + "multiqc_versions_config": { + "content": [ + [ + "versions.yml:md5,21f35ee29416b9b3073c28733efe4b7d" + ] + ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.10.1" + }, + "timestamp": "2024-02-29T08:49:25.457567" + } +} \ No newline at end of file diff --git a/modules/nf-core/multiqc/tests/tags.yml b/modules/nf-core/multiqc/tests/tags.yml new file mode 100644 index 00000000..bea6c0d3 --- /dev/null +++ b/modules/nf-core/multiqc/tests/tags.yml @@ -0,0 +1,2 @@ +multiqc: + - modules/nf-core/multiqc/** diff --git a/nextflow.config b/nextflow.config new file mode 100644 index 00000000..b9c27d9e --- /dev/null +++ b/nextflow.config @@ -0,0 +1,280 @@ +/* +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ + nf-core/rnadeseq Nextflow config file +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ + Default config options for all compute environments +---------------------------------------------------------------------------------------- +*/ + +// Global default params, used in configs +params { + + // TODO nf-core: Specify your pipeline's command line flags + // Input options + input = null + // References + genome = null + igenomes_base = 's3://ngi-igenomes/igenomes/' + igenomes_ignore = false + + // MultiQC options + multiqc_config = null + multiqc_title = null + multiqc_logo = null + max_multiqc_email_size = '25.MB' + multiqc_methods_description = null + + // Boilerplate options + outdir = null + publish_dir_mode = 'copy' + email = null + email_on_fail = null + plaintext_email = false + monochrome_logs = false + hook_url = null + help = false + version = false + pipelines_testdata_base_path = 'https://raw.githubusercontent.com/nf-core/test-datasets/' + + // Config options + config_profile_name = null + config_profile_description = null + custom_config_version = 'master' + custom_config_base = "https://raw.githubusercontent.com/nf-core/configs/${params.custom_config_version}" + config_profile_contact = null + config_profile_url = null + + // Max resource options + // Defaults only, expecting to be overwritten + max_memory = '128.GB' + max_cpus = 16 + max_time = '240.h' + + // Schema validation default options + validationFailUnrecognisedParams = false + validationLenientMode = false + validationSchemaIgnoreParams = 'genomes,igenomes_base' + validationShowHiddenParams = false + validate_params = true + +} + +// Load base.config by default for all pipelines +includeConfig 'conf/base.config' + +// Load nf-core custom profiles from different Institutions +try { + includeConfig "${params.custom_config_base}/nfcore_custom.config" +} catch (Exception e) { + System.err.println("WARNING: Could not load nf-core/config profiles: ${params.custom_config_base}/nfcore_custom.config") +} + +// Load nf-core/rnadeseq custom profiles from different institutions. +try { + includeConfig "${params.custom_config_base}/pipeline/rnadeseq.config" +} catch (Exception e) { + System.err.println("WARNING: Could not load nf-core/config/rnadeseq profiles: ${params.custom_config_base}/pipeline/rnadeseq.config") +} +profiles { + debug { + dumpHashes = true + process.beforeScript = 'echo $HOSTNAME' + cleanup = false + nextflow.enable.configProcessNamesValidation = true + } + conda { + conda.enabled = true + docker.enabled = false + singularity.enabled = false + podman.enabled = false + shifter.enabled = false + charliecloud.enabled = false + conda.channels = ['conda-forge', 'bioconda', 'defaults'] + apptainer.enabled = false + } + mamba { + conda.enabled = true + conda.useMamba = true + docker.enabled = false + singularity.enabled = false + podman.enabled = false + shifter.enabled = false + charliecloud.enabled = false + apptainer.enabled = false + } + docker { + docker.enabled = true + conda.enabled = false + singularity.enabled = false + podman.enabled = false + shifter.enabled = false + charliecloud.enabled = false + apptainer.enabled = false + docker.runOptions = '-u $(id -u):$(id -g)' + } + arm { + docker.runOptions = '-u $(id -u):$(id -g) --platform=linux/amd64' + } + singularity { + singularity.enabled = true + singularity.autoMounts = true + conda.enabled = false + docker.enabled = false + podman.enabled = false + shifter.enabled = false + charliecloud.enabled = false + apptainer.enabled = false + } + podman { + podman.enabled = true + conda.enabled = false + docker.enabled = false + singularity.enabled = false + shifter.enabled = false + charliecloud.enabled = false + apptainer.enabled = false + } + shifter { + shifter.enabled = true + conda.enabled = false + docker.enabled = false + singularity.enabled = false + podman.enabled = false + charliecloud.enabled = false + apptainer.enabled = false + } + charliecloud { + charliecloud.enabled = true + conda.enabled = false + docker.enabled = false + singularity.enabled = false + podman.enabled = false + shifter.enabled = false + apptainer.enabled = false + } + apptainer { + apptainer.enabled = true + apptainer.autoMounts = true + conda.enabled = false + docker.enabled = false + singularity.enabled = false + podman.enabled = false + shifter.enabled = false + charliecloud.enabled = false + } + wave { + apptainer.ociAutoPull = true + singularity.ociAutoPull = true + wave.enabled = true + wave.freeze = true + wave.strategy = 'conda,container' + } + gitpod { + executor.name = 'local' + executor.cpus = 4 + executor.memory = 8.GB + } + test { includeConfig 'conf/test.config' } + test_full { includeConfig 'conf/test_full.config' } +} + +// Set default registry for Apptainer, Docker, Podman and Singularity independent of -profile +// Will not be used unless Apptainer / Docker / Podman / Singularity are enabled +// Set to your registry if you have a mirror of containers +apptainer.registry = 'quay.io' +docker.registry = 'quay.io' +podman.registry = 'quay.io' +singularity.registry = 'quay.io' + +// Nextflow plugins +plugins { + id 'nf-validation@1.1.3' // Validation of pipeline parameters and creation of an input channel from a sample sheet +} + +// Load igenomes.config if required +if (!params.igenomes_ignore) { + includeConfig 'conf/igenomes.config' +} else { + params.genomes = [:] +} +// Export these variables to prevent local Python/R libraries from conflicting with those in the container +// The JULIA depot path has been adjusted to a fixed path `/usr/local/share/julia` that needs to be used for packages in the container. +// See https://apeltzer.github.io/post/03-julia-lang-nextflow/ for details on that. Once we have a common agreement on where to keep Julia packages, this is adjustable. + +env { + PYTHONNOUSERSITE = 1 + R_PROFILE_USER = "/.Rprofile" + R_ENVIRON_USER = "/.Renviron" + JULIA_DEPOT_PATH = "/usr/local/share/julia" +} + +// Capture exit codes from upstream processes when piping +process.shell = ['/bin/bash', '-euo', 'pipefail'] + +// Disable process selector warnings by default. Use debug profile to enable warnings. +nextflow.enable.configProcessNamesValidation = false + +def trace_timestamp = new java.util.Date().format( 'yyyy-MM-dd_HH-mm-ss') +timeline { + enabled = true + file = "${params.outdir}/pipeline_info/execution_timeline_${trace_timestamp}.html" +} +report { + enabled = true + file = "${params.outdir}/pipeline_info/execution_report_${trace_timestamp}.html" +} +trace { + enabled = true + file = "${params.outdir}/pipeline_info/execution_trace_${trace_timestamp}.txt" +} +dag { + enabled = true + file = "${params.outdir}/pipeline_info/pipeline_dag_${trace_timestamp}.html" +} + +manifest { + name = 'nf-core/rnadeseq' + author = """""" + homePage = 'https://github.com/nf-core/rnadeseq' + description = """""" + mainScript = 'main.nf' + nextflowVersion = '!>=23.04.0' + version = '1.0dev' + doi = '' +} + +// Load modules.config for DSL2 module specific options +includeConfig 'conf/modules.config' + +// Function to ensure that resource requirements don't go beyond +// a maximum limit +def check_max(obj, type) { + if (type == 'memory') { + try { + if (obj.compareTo(params.max_memory as nextflow.util.MemoryUnit) == 1) + return params.max_memory as nextflow.util.MemoryUnit + else + return obj + } catch (all) { + println " ### ERROR ### Max memory '${params.max_memory}' is not valid! Using default value: $obj" + return obj + } + } else if (type == 'time') { + try { + if (obj.compareTo(params.max_time as nextflow.util.Duration) == 1) + return params.max_time as nextflow.util.Duration + else + return obj + } catch (all) { + println " ### ERROR ### Max time '${params.max_time}' is not valid! Using default value: $obj" + return obj + } + } else if (type == 'cpus') { + try { + return Math.min( obj, params.max_cpus as int ) + } catch (all) { + println " ### ERROR ### Max cpus '${params.max_cpus}' is not valid! Using default value: $obj" + return obj + } + } +} diff --git a/nextflow_schema.json b/nextflow_schema.json new file mode 100644 index 00000000..4e3b45b0 --- /dev/null +++ b/nextflow_schema.json @@ -0,0 +1,296 @@ +{ + "$schema": "http://json-schema.org/draft-07/schema", + "$id": "https://raw.githubusercontent.com/nf-core/rnadeseq/master/nextflow_schema.json", + "title": "nf-core/rnadeseq pipeline parameters", + "description": "", + "type": "object", + "definitions": { + "input_output_options": { + "title": "Input/output options", + "type": "object", + "fa_icon": "fas fa-terminal", + "description": "Define where the pipeline should find input data and save output data.", + "required": ["input", "outdir"], + "properties": { + "input": { + "type": "string", + "format": "file-path", + "exists": true, + "schema": "assets/schema_input.json", + "mimetype": "text/csv", + "pattern": "^\\S+\\.csv$", + "description": "Path to comma-separated file containing information about the samples in the experiment.", + "help_text": "You will need to create a design file with information about the samples in your experiment before running the pipeline. Use this parameter to specify its location. It has to be a comma-separated file with 3 columns, and a header row. See [usage docs](https://nf-co.re/rnadeseq/usage#samplesheet-input).", + "fa_icon": "fas fa-file-csv" + }, + "outdir": { + "type": "string", + "format": "directory-path", + "description": "The output directory where the results will be saved. You have to use absolute paths to storage on Cloud infrastructure.", + "fa_icon": "fas fa-folder-open" + }, + "email": { + "type": "string", + "description": "Email address for completion summary.", + "fa_icon": "fas fa-envelope", + "help_text": "Set this parameter to your e-mail address to get a summary e-mail with details of the run sent to you when the workflow exits. If set in your user config file (`~/.nextflow/config`) then you don't need to specify this on the command line for every run.", + "pattern": "^([a-zA-Z0-9_\\-\\.]+)@([a-zA-Z0-9_\\-\\.]+)\\.([a-zA-Z]{2,5})$" + }, + "multiqc_title": { + "type": "string", + "description": "MultiQC report title. Printed as page header, used for filename if not otherwise specified.", + "fa_icon": "fas fa-file-signature" + } + } + }, + "reference_genome_options": { + "title": "Reference genome options", + "type": "object", + "fa_icon": "fas fa-dna", + "description": "Reference genome related files and options required for the workflow.", + "properties": { + "genome": { + "type": "string", + "description": "Name of iGenomes reference.", + "fa_icon": "fas fa-book", + "help_text": "If using a reference genome configured in the pipeline using iGenomes, use this parameter to give the ID for the reference. This is then used to build the full paths for all required reference genome files e.g. `--genome GRCh38`. \n\nSee the [nf-core website docs](https://nf-co.re/usage/reference_genomes) for more details." + }, + "fasta": { + "type": "string", + "format": "file-path", + "exists": true, + "mimetype": "text/plain", + "pattern": "^\\S+\\.fn?a(sta)?(\\.gz)?$", + "description": "Path to FASTA genome file.", + "help_text": "This parameter is *mandatory* if `--genome` is not specified. If you don't have a BWA index available this will be generated for you automatically. Combine with `--save_reference` to save BWA index for future runs.", + "fa_icon": "far fa-file-code" + }, + "igenomes_ignore": { + "type": "boolean", + "description": "Do not load the iGenomes reference config.", + "fa_icon": "fas fa-ban", + "hidden": true, + "help_text": "Do not load `igenomes.config` when running the pipeline. You may choose this option if you observe clashes between custom parameters and those supplied in `igenomes.config`." + } + } + }, + "institutional_config_options": { + "title": "Institutional config options", + "type": "object", + "fa_icon": "fas fa-university", + "description": "Parameters used to describe centralised config profiles. These should not be edited.", + "help_text": "The centralised nf-core configuration profiles use a handful of pipeline parameters to describe themselves. This information is then printed to the Nextflow log when you run a pipeline. You should not need to change these values when you run a pipeline.", + "properties": { + "custom_config_version": { + "type": "string", + "description": "Git commit id for Institutional configs.", + "default": "master", + "hidden": true, + "fa_icon": "fas fa-users-cog" + }, + "custom_config_base": { + "type": "string", + "description": "Base directory for Institutional configs.", + "default": "https://raw.githubusercontent.com/nf-core/configs/master", + "hidden": true, + "help_text": "If you're running offline, Nextflow will not be able to fetch the institutional config files from the internet. If you don't need them, then this is not a problem. If you do need them, you should download the files from the repo and tell Nextflow where to find them with this parameter.", + "fa_icon": "fas fa-users-cog" + }, + "config_profile_name": { + "type": "string", + "description": "Institutional config name.", + "hidden": true, + "fa_icon": "fas fa-users-cog" + }, + "config_profile_description": { + "type": "string", + "description": "Institutional config description.", + "hidden": true, + "fa_icon": "fas fa-users-cog" + }, + "config_profile_contact": { + "type": "string", + "description": "Institutional config contact information.", + "hidden": true, + "fa_icon": "fas fa-users-cog" + }, + "config_profile_url": { + "type": "string", + "description": "Institutional config URL link.", + "hidden": true, + "fa_icon": "fas fa-users-cog" + } + } + }, + "max_job_request_options": { + "title": "Max job request options", + "type": "object", + "fa_icon": "fab fa-acquisitions-incorporated", + "description": "Set the top limit for requested resources for any single job.", + "help_text": "If you are running on a smaller system, a pipeline step requesting more resources than are available may cause the Nextflow to stop the run with an error. These options allow you to cap the maximum resources requested by any single job so that the pipeline will run on your system.\n\nNote that you can not _increase_ the resources requested by any job using these options. For that you will need your own configuration file. See [the nf-core website](https://nf-co.re/usage/configuration) for details.", + "properties": { + "max_cpus": { + "type": "integer", + "description": "Maximum number of CPUs that can be requested for any single job.", + "default": 16, + "fa_icon": "fas fa-microchip", + "hidden": true, + "help_text": "Use to set an upper-limit for the CPU requirement for each process. Should be an integer e.g. `--max_cpus 1`" + }, + "max_memory": { + "type": "string", + "description": "Maximum amount of memory that can be requested for any single job.", + "default": "128.GB", + "fa_icon": "fas fa-memory", + "pattern": "^\\d+(\\.\\d+)?\\.?\\s*(K|M|G|T)?B$", + "hidden": true, + "help_text": "Use to set an upper-limit for the memory requirement for each process. Should be a string in the format integer-unit e.g. `--max_memory '8.GB'`" + }, + "max_time": { + "type": "string", + "description": "Maximum amount of time that can be requested for any single job.", + "default": "240.h", + "fa_icon": "far fa-clock", + "pattern": "^(\\d+\\.?\\s*(s|m|h|d|day)\\s*)+$", + "hidden": true, + "help_text": "Use to set an upper-limit for the time requirement for each process. Should be a string in the format integer-unit e.g. `--max_time '2.h'`" + } + } + }, + "generic_options": { + "title": "Generic options", + "type": "object", + "fa_icon": "fas fa-file-import", + "description": "Less common options for the pipeline, typically set in a config file.", + "help_text": "These options are common to all nf-core pipelines and allow you to customise some of the core preferences for how the pipeline runs.\n\nTypically these options would be set in a Nextflow config file loaded for all pipeline runs, such as `~/.nextflow/config`.", + "properties": { + "help": { + "type": "boolean", + "description": "Display help text.", + "fa_icon": "fas fa-question-circle", + "hidden": true + }, + "version": { + "type": "boolean", + "description": "Display version and exit.", + "fa_icon": "fas fa-question-circle", + "hidden": true + }, + "publish_dir_mode": { + "type": "string", + "default": "copy", + "description": "Method used to save pipeline results to output directory.", + "help_text": "The Nextflow `publishDir` option specifies which intermediate files should be saved to the output directory. This option tells the pipeline what method should be used to move these files. See [Nextflow docs](https://www.nextflow.io/docs/latest/process.html#publishdir) for details.", + "fa_icon": "fas fa-copy", + "enum": ["symlink", "rellink", "link", "copy", "copyNoFollow", "move"], + "hidden": true + }, + "email_on_fail": { + "type": "string", + "description": "Email address for completion summary, only when pipeline fails.", + "fa_icon": "fas fa-exclamation-triangle", + "pattern": "^([a-zA-Z0-9_\\-\\.]+)@([a-zA-Z0-9_\\-\\.]+)\\.([a-zA-Z]{2,5})$", + "help_text": "An email address to send a summary email to when the pipeline is completed - ONLY sent if the pipeline does not exit successfully.", + "hidden": true + }, + "plaintext_email": { + "type": "boolean", + "description": "Send plain-text email instead of HTML.", + "fa_icon": "fas fa-remove-format", + "hidden": true + }, + "max_multiqc_email_size": { + "type": "string", + "description": "File size limit when attaching MultiQC reports to summary emails.", + "pattern": "^\\d+(\\.\\d+)?\\.?\\s*(K|M|G|T)?B$", + "default": "25.MB", + "fa_icon": "fas fa-file-upload", + "hidden": true + }, + "monochrome_logs": { + "type": "boolean", + "description": "Do not use coloured log outputs.", + "fa_icon": "fas fa-palette", + "hidden": true + }, + "hook_url": { + "type": "string", + "description": "Incoming hook URL for messaging service", + "fa_icon": "fas fa-people-group", + "help_text": "Incoming hook URL for messaging service. Currently, MS Teams and Slack are supported.", + "hidden": true + }, + "multiqc_config": { + "type": "string", + "format": "file-path", + "description": "Custom config file to supply to MultiQC.", + "fa_icon": "fas fa-cog", + "hidden": true + }, + "multiqc_logo": { + "type": "string", + "description": "Custom logo file to supply to MultiQC. File name must also be set in the MultiQC config file", + "fa_icon": "fas fa-image", + "hidden": true + }, + "multiqc_methods_description": { + "type": "string", + "description": "Custom MultiQC yaml file containing HTML including a methods description.", + "fa_icon": "fas fa-cog" + }, + "validate_params": { + "type": "boolean", + "description": "Boolean whether to validate parameters against the schema at runtime", + "default": true, + "fa_icon": "fas fa-check-square", + "hidden": true + }, + "validationShowHiddenParams": { + "type": "boolean", + "fa_icon": "far fa-eye-slash", + "description": "Show all params when using `--help`", + "hidden": true, + "help_text": "By default, parameters set as _hidden_ in the schema are not shown on the command line when a user runs with `--help`. Specifying this option will tell the pipeline to show all parameters." + }, + "validationFailUnrecognisedParams": { + "type": "boolean", + "fa_icon": "far fa-check-circle", + "description": "Validation of parameters fails when an unrecognised parameter is found.", + "hidden": true, + "help_text": "By default, when an unrecognised parameter is found, it returns a warinig." + }, + "validationLenientMode": { + "type": "boolean", + "fa_icon": "far fa-check-circle", + "description": "Validation of parameters in lenient more.", + "hidden": true, + "help_text": "Allows string values that are parseable as numbers or booleans. For further information see [JSONSchema docs](https://github.com/everit-org/json-schema#lenient-mode)." + }, + "pipelines_testdata_base_path": { + "type": "string", + "fa_icon": "far fa-check-circle", + "description": "Base URL or local path to location of pipeline test dataset files", + "default": "https://raw.githubusercontent.com/nf-core/test-datasets/", + "hidden": true + } + } + } + }, + "allOf": [ + { + "$ref": "#/definitions/input_output_options" + }, + { + "$ref": "#/definitions/reference_genome_options" + }, + { + "$ref": "#/definitions/institutional_config_options" + }, + { + "$ref": "#/definitions/max_job_request_options" + }, + { + "$ref": "#/definitions/generic_options" + } + ] +} diff --git a/subworkflows/local/utils_nfcore_rnadeseq_pipeline/main.nf b/subworkflows/local/utils_nfcore_rnadeseq_pipeline/main.nf new file mode 100644 index 00000000..850d62cc --- /dev/null +++ b/subworkflows/local/utils_nfcore_rnadeseq_pipeline/main.nf @@ -0,0 +1,263 @@ +// +// Subworkflow with functionality specific to the nf-core/rnadeseq pipeline +// + +/* +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ + IMPORT FUNCTIONS / MODULES / SUBWORKFLOWS +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ +*/ + +include { UTILS_NFVALIDATION_PLUGIN } from '../../nf-core/utils_nfvalidation_plugin' +include { paramsSummaryMap } from 'plugin/nf-validation' +include { fromSamplesheet } from 'plugin/nf-validation' +include { UTILS_NEXTFLOW_PIPELINE } from '../../nf-core/utils_nextflow_pipeline' +include { completionEmail } from '../../nf-core/utils_nfcore_pipeline' +include { completionSummary } from '../../nf-core/utils_nfcore_pipeline' +include { dashedLine } from '../../nf-core/utils_nfcore_pipeline' +include { nfCoreLogo } from '../../nf-core/utils_nfcore_pipeline' +include { imNotification } from '../../nf-core/utils_nfcore_pipeline' +include { UTILS_NFCORE_PIPELINE } from '../../nf-core/utils_nfcore_pipeline' +include { workflowCitation } from '../../nf-core/utils_nfcore_pipeline' + +/* +======================================================================================== + SUBWORKFLOW TO INITIALISE PIPELINE +======================================================================================== +*/ + +workflow PIPELINE_INITIALISATION { + + take: + version // boolean: Display version and exit + help // boolean: Display help text + validate_params // boolean: Boolean whether to validate parameters against the schema at runtime + monochrome_logs // boolean: Do not use coloured log outputs + nextflow_cli_args // array: List of positional nextflow CLI args + outdir // string: The output directory where the results will be saved + input // string: Path to input samplesheet + + main: + + ch_versions = Channel.empty() + + // + // Print version and exit if required and dump pipeline parameters to JSON file + // + UTILS_NEXTFLOW_PIPELINE ( + version, + true, + outdir, + workflow.profile.tokenize(',').intersect(['conda', 'mamba']).size() >= 1 + ) + + // + // Validate parameters and generate parameter summary to stdout + // + pre_help_text = nfCoreLogo(monochrome_logs) + post_help_text = '\n' + workflowCitation() + '\n' + dashedLine(monochrome_logs) + def String workflow_command = "nextflow run ${workflow.manifest.name} -profile --input samplesheet.csv --outdir " + UTILS_NFVALIDATION_PLUGIN ( + help, + workflow_command, + pre_help_text, + post_help_text, + validate_params, + "nextflow_schema.json" + ) + + // + // Check config provided to the pipeline + // + UTILS_NFCORE_PIPELINE ( + nextflow_cli_args + ) + // + // Custom validation for pipeline parameters + // + validateInputParameters() + + // + // Create channel from input file provided through params.input + // + Channel + .fromSamplesheet("input") + .map { + meta, fastq_1, fastq_2 -> + if (!fastq_2) { + return [ meta.id, meta + [ single_end:true ], [ fastq_1 ] ] + } else { + return [ meta.id, meta + [ single_end:false ], [ fastq_1, fastq_2 ] ] + } + } + .groupTuple() + .map { + validateInputSamplesheet(it) + } + .map { + meta, fastqs -> + return [ meta, fastqs.flatten() ] + } + .set { ch_samplesheet } + + emit: + samplesheet = ch_samplesheet + versions = ch_versions +} + +/* +======================================================================================== + SUBWORKFLOW FOR PIPELINE COMPLETION +======================================================================================== +*/ + +workflow PIPELINE_COMPLETION { + + take: + email // string: email address + email_on_fail // string: email address sent on pipeline failure + plaintext_email // boolean: Send plain-text email instead of HTML + outdir // path: Path to output directory where results will be published + monochrome_logs // boolean: Disable ANSI colour codes in log output + hook_url // string: hook URL for notifications + multiqc_report // string: Path to MultiQC report + + main: + + summary_params = paramsSummaryMap(workflow, parameters_schema: "nextflow_schema.json") + + // + // Completion email and summary + // + workflow.onComplete { + if (email || email_on_fail) { + completionEmail(summary_params, email, email_on_fail, plaintext_email, outdir, monochrome_logs, multiqc_report.toList()) + } + + completionSummary(monochrome_logs) + + if (hook_url) { + imNotification(summary_params, hook_url) + } + } + + workflow.onError { + log.error "Pipeline failed. Please refer to troubleshooting docs: https://nf-co.re/docs/usage/troubleshooting" + } +} + +/* +======================================================================================== + FUNCTIONS +======================================================================================== +*/ +// +// Check and validate pipeline parameters +// +def validateInputParameters() { + genomeExistsError() +} + +// +// Validate channels from input samplesheet +// +def validateInputSamplesheet(input) { + def (metas, fastqs) = input[1..2] + + // Check that multiple runs of the same sample are of the same datatype i.e. single-end / paired-end + def endedness_ok = metas.collect{ it.single_end }.unique().size == 1 + if (!endedness_ok) { + error("Please check input samplesheet -> Multiple runs of a sample must be of the same datatype i.e. single-end or paired-end: ${metas[0].id}") + } + + return [ metas[0], fastqs ] +} +// +// Get attribute from genome config file e.g. fasta +// +def getGenomeAttribute(attribute) { + if (params.genomes && params.genome && params.genomes.containsKey(params.genome)) { + if (params.genomes[ params.genome ].containsKey(attribute)) { + return params.genomes[ params.genome ][ attribute ] + } + } + return null +} + +// +// Exit pipeline if incorrect --genome key provided +// +def genomeExistsError() { + if (params.genomes && params.genome && !params.genomes.containsKey(params.genome)) { + def error_string = "~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~\n" + + " Genome '${params.genome}' not found in any config files provided to the pipeline.\n" + + " Currently, the available genome keys are:\n" + + " ${params.genomes.keySet().join(", ")}\n" + + "~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~" + error(error_string) + } +} + +// +// Generate methods description for MultiQC +// +def toolCitationText() { + // TODO nf-core: Optionally add in-text citation tools to this list. + // Can use ternary operators to dynamically construct based conditions, e.g. params["run_xyz"] ? "Tool (Foo et al. 2023)" : "", + // Uncomment function in methodsDescriptionText to render in MultiQC report + def citation_text = [ + "Tools used in the workflow included:", + "FastQC (Andrews 2010),", + "MultiQC (Ewels et al. 2016)", + "." + ].join(' ').trim() + + return citation_text +} + +def toolBibliographyText() { + // TODO nf-core: Optionally add bibliographic entries to this list. + // Can use ternary operators to dynamically construct based conditions, e.g. params["run_xyz"] ? "
  • Author (2023) Pub name, Journal, DOI
    • " : "", + // Uncomment function in methodsDescriptionText to render in MultiQC report + def reference_text = [ + "
  • Andrews S, (2010) FastQC, URL: https://www.bioinformatics.babraham.ac.uk/projects/fastqc/).
    • ", + "
  • Ewels, P., Magnusson, M., Lundin, S., & Käller, M. (2016). MultiQC: summarize analysis results for multiple tools and samples in a single report. Bioinformatics , 32(19), 3047–3048. doi: /10.1093/bioinformatics/btw354
    • " + ].join(' ').trim() + + return reference_text +} + +def methodsDescriptionText(mqc_methods_yaml) { + // Convert to a named map so can be used as with familar NXF ${workflow} variable syntax in the MultiQC YML file + def meta = [:] + meta.workflow = workflow.toMap() + meta["manifest_map"] = workflow.manifest.toMap() + + // Pipeline DOI + if (meta.manifest_map.doi) { + // Using a loop to handle multiple DOIs + // Removing `https://doi.org/` to handle pipelines using DOIs vs DOI resolvers + // Removing ` ` since the manifest.doi is a string and not a proper list + def temp_doi_ref = "" + String[] manifest_doi = meta.manifest_map.doi.tokenize(",") + for (String doi_ref: manifest_doi) temp_doi_ref += "(doi:     ${doi_ref.replace("https://doi.org/", "").replace(" ", "")}), " + meta["doi_text"] = temp_doi_ref.substring(0, temp_doi_ref.length() - 2) + } else meta["doi_text"] = "" + meta["nodoi_text"] = meta.manifest_map.doi ? "" : "
  • If available, make sure to update the text to include the Zenodo DOI of version of the pipeline used.
    • " + + // Tool references + meta["tool_citations"] = "" + meta["tool_bibliography"] = "" + + // TODO nf-core: Only uncomment below if logic in toolCitationText/toolBibliographyText has been filled! + // meta["tool_citations"] = toolCitationText().replaceAll(", \\.", ".").replaceAll("\\. \\.", ".").replaceAll(", \\.", ".") + // meta["tool_bibliography"] = toolBibliographyText() + + + def methods_text = mqc_methods_yaml.text + + def engine = new groovy.text.SimpleTemplateEngine() + def description_html = engine.createTemplate(methods_text).make(meta) + + return description_html.toString() +} diff --git a/subworkflows/nf-core/utils_nextflow_pipeline/main.nf b/subworkflows/nf-core/utils_nextflow_pipeline/main.nf new file mode 100644 index 00000000..ac31f28f --- /dev/null +++ b/subworkflows/nf-core/utils_nextflow_pipeline/main.nf @@ -0,0 +1,126 @@ +// +// Subworkflow with functionality that may be useful for any Nextflow pipeline +// + +import org.yaml.snakeyaml.Yaml +import groovy.json.JsonOutput +import nextflow.extension.FilesEx + +/* +======================================================================================== + SUBWORKFLOW DEFINITION +======================================================================================== +*/ + +workflow UTILS_NEXTFLOW_PIPELINE { + + take: + print_version // boolean: print version + dump_parameters // boolean: dump parameters + outdir // path: base directory used to publish pipeline results + check_conda_channels // boolean: check conda channels + + main: + + // + // Print workflow version and exit on --version + // + if (print_version) { + log.info "${workflow.manifest.name} ${getWorkflowVersion()}" + System.exit(0) + } + + // + // Dump pipeline parameters to a JSON file + // + if (dump_parameters && outdir) { + dumpParametersToJSON(outdir) + } + + // + // When running with Conda, warn if channels have not been set-up appropriately + // + if (check_conda_channels) { + checkCondaChannels() + } + + emit: + dummy_emit = true +} + +/* +======================================================================================== + FUNCTIONS +======================================================================================== +*/ + +// +// Generate version string +// +def getWorkflowVersion() { + String version_string = "" + if (workflow.manifest.version) { + def prefix_v = workflow.manifest.version[0] != 'v' ? 'v' : '' + version_string += "${prefix_v}${workflow.manifest.version}" + } + + if (workflow.commitId) { + def git_shortsha = workflow.commitId.substring(0, 7) + version_string += "-g${git_shortsha}" + } + + return version_string +} + +// +// Dump pipeline parameters to a JSON file +// +def dumpParametersToJSON(outdir) { + def timestamp = new java.util.Date().format( 'yyyy-MM-dd_HH-mm-ss') + def filename = "params_${timestamp}.json" + def temp_pf = new File(workflow.launchDir.toString(), ".${filename}") + def jsonStr = JsonOutput.toJson(params) + temp_pf.text = JsonOutput.prettyPrint(jsonStr) + + FilesEx.copyTo(temp_pf.toPath(), "${outdir}/pipeline_info/params_${timestamp}.json") + temp_pf.delete() +} + +// +// When running with -profile conda, warn if channels have not been set-up appropriately +// +def checkCondaChannels() { + Yaml parser = new Yaml() + def channels = [] + try { + def config = parser.load("conda config --show channels".execute().text) + channels = config.channels + } catch(NullPointerException | IOException e) { + log.warn "Could not verify conda channel configuration." + return + } + + // Check that all channels are present + // This channel list is ordered by required channel priority. + def required_channels_in_order = ['conda-forge', 'bioconda', 'defaults'] + def channels_missing = ((required_channels_in_order as Set) - (channels as Set)) as Boolean + + // Check that they are in the right order + def channel_priority_violation = false + def n = required_channels_in_order.size() + for (int i = 0; i < n - 1; i++) { + channel_priority_violation |= !(channels.indexOf(required_channels_in_order[i]) < channels.indexOf(required_channels_in_order[i+1])) + } + + if (channels_missing | channel_priority_violation) { + log.warn "~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~\n" + + " There is a problem with your Conda configuration!\n\n" + + " You will need to set-up the conda-forge and bioconda channels correctly.\n" + + " Please refer to https://bioconda.github.io/\n" + + " The observed channel order is \n" + + " ${channels}\n" + + " but the following channel order is required:\n" + + " ${required_channels_in_order}\n" + + "~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~" + } +} diff --git a/subworkflows/nf-core/utils_nextflow_pipeline/meta.yml b/subworkflows/nf-core/utils_nextflow_pipeline/meta.yml new file mode 100644 index 00000000..e5c3a0a8 --- /dev/null +++ b/subworkflows/nf-core/utils_nextflow_pipeline/meta.yml @@ -0,0 +1,38 @@ +# yaml-language-server: $schema=https://raw.githubusercontent.com/nf-core/modules/master/subworkflows/yaml-schema.json +name: "UTILS_NEXTFLOW_PIPELINE" +description: Subworkflow with functionality that may be useful for any Nextflow pipeline +keywords: + - utility + - pipeline + - initialise + - version +components: [] +input: + - print_version: + type: boolean + description: | + Print the version of the pipeline and exit + - dump_parameters: + type: boolean + description: | + Dump the parameters of the pipeline to a JSON file + - output_directory: + type: directory + description: Path to output dir to write JSON file to. + pattern: "results/" + - check_conda_channel: + type: boolean + description: | + Check if the conda channel priority is correct. +output: + - dummy_emit: + type: boolean + description: | + Dummy emit to make nf-core subworkflows lint happy +authors: + - "@adamrtalbot" + - "@drpatelh" +maintainers: + - "@adamrtalbot" + - "@drpatelh" + - "@maxulysse" diff --git a/subworkflows/nf-core/utils_nextflow_pipeline/tests/main.function.nf.test b/subworkflows/nf-core/utils_nextflow_pipeline/tests/main.function.nf.test new file mode 100644 index 00000000..68718e4f --- /dev/null +++ b/subworkflows/nf-core/utils_nextflow_pipeline/tests/main.function.nf.test @@ -0,0 +1,54 @@ + +nextflow_function { + + name "Test Functions" + script "subworkflows/nf-core/utils_nextflow_pipeline/main.nf" + config "subworkflows/nf-core/utils_nextflow_pipeline/tests/nextflow.config" + tag 'subworkflows' + tag 'utils_nextflow_pipeline' + tag 'subworkflows/utils_nextflow_pipeline' + + test("Test Function getWorkflowVersion") { + + function "getWorkflowVersion" + + then { + assertAll( + { assert function.success }, + { assert snapshot(function.result).match() } + ) + } + } + + test("Test Function dumpParametersToJSON") { + + function "dumpParametersToJSON" + + when { + function { + """ + // define inputs of the function here. Example: + input[0] = "$outputDir" + """.stripIndent() + } + } + + then { + assertAll( + { assert function.success } + ) + } + } + + test("Test Function checkCondaChannels") { + + function "checkCondaChannels" + + then { + assertAll( + { assert function.success }, + { assert snapshot(function.result).match() } + ) + } + } +} diff --git a/subworkflows/nf-core/utils_nextflow_pipeline/tests/main.function.nf.test.snap b/subworkflows/nf-core/utils_nextflow_pipeline/tests/main.function.nf.test.snap new file mode 100644 index 00000000..e3f0baf4 --- /dev/null +++ b/subworkflows/nf-core/utils_nextflow_pipeline/tests/main.function.nf.test.snap @@ -0,0 +1,20 @@ +{ + "Test Function getWorkflowVersion": { + "content": [ + "v9.9.9" + ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.10.1" + }, + "timestamp": "2024-02-28T12:02:05.308243" + }, + "Test Function checkCondaChannels": { + "content": null, + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.10.1" + }, + "timestamp": "2024-02-28T12:02:12.425833" + } +} \ No newline at end of file diff --git a/subworkflows/nf-core/utils_nextflow_pipeline/tests/main.workflow.nf.test b/subworkflows/nf-core/utils_nextflow_pipeline/tests/main.workflow.nf.test new file mode 100644 index 00000000..ca964ce8 --- /dev/null +++ b/subworkflows/nf-core/utils_nextflow_pipeline/tests/main.workflow.nf.test @@ -0,0 +1,111 @@ +nextflow_workflow { + + name "Test Workflow UTILS_NEXTFLOW_PIPELINE" + script "../main.nf" + config "subworkflows/nf-core/utils_nextflow_pipeline/tests/nextflow.config" + workflow "UTILS_NEXTFLOW_PIPELINE" + tag 'subworkflows' + tag 'utils_nextflow_pipeline' + tag 'subworkflows/utils_nextflow_pipeline' + + test("Should run no inputs") { + + when { + workflow { + """ + print_version = false + dump_parameters = false + outdir = null + check_conda_channels = false + + input[0] = print_version + input[1] = dump_parameters + input[2] = outdir + input[3] = check_conda_channels + """ + } + } + + then { + assertAll( + { assert workflow.success } + ) + } + } + + test("Should print version") { + + when { + workflow { + """ + print_version = true + dump_parameters = false + outdir = null + check_conda_channels = false + + input[0] = print_version + input[1] = dump_parameters + input[2] = outdir + input[3] = check_conda_channels + """ + } + } + + then { + assertAll( + { assert workflow.success }, + { assert workflow.stdout.contains("nextflow_workflow v9.9.9") } + ) + } + } + + test("Should dump params") { + + when { + workflow { + """ + print_version = false + dump_parameters = true + outdir = 'results' + check_conda_channels = false + + input[0] = false + input[1] = true + input[2] = outdir + input[3] = false + """ + } + } + + then { + assertAll( + { assert workflow.success } + ) + } + } + + test("Should not create params JSON if no output directory") { + + when { + workflow { + """ + print_version = false + dump_parameters = true + outdir = null + check_conda_channels = false + + input[0] = false + input[1] = true + input[2] = outdir + input[3] = false + """ + } + } + + then { + assertAll( + { assert workflow.success } + ) + } + } +} diff --git a/subworkflows/nf-core/utils_nextflow_pipeline/tests/nextflow.config b/subworkflows/nf-core/utils_nextflow_pipeline/tests/nextflow.config new file mode 100644 index 00000000..d0a926bf --- /dev/null +++ b/subworkflows/nf-core/utils_nextflow_pipeline/tests/nextflow.config @@ -0,0 +1,9 @@ +manifest { + name = 'nextflow_workflow' + author = """nf-core""" + homePage = 'https://127.0.0.1' + description = """Dummy pipeline""" + nextflowVersion = '!>=23.04.0' + version = '9.9.9' + doi = 'https://doi.org/10.5281/zenodo.5070524' +} diff --git a/subworkflows/nf-core/utils_nextflow_pipeline/tests/tags.yml b/subworkflows/nf-core/utils_nextflow_pipeline/tests/tags.yml new file mode 100644 index 00000000..f8476112 --- /dev/null +++ b/subworkflows/nf-core/utils_nextflow_pipeline/tests/tags.yml @@ -0,0 +1,2 @@ +subworkflows/utils_nextflow_pipeline: + - subworkflows/nf-core/utils_nextflow_pipeline/** diff --git a/subworkflows/nf-core/utils_nfcore_pipeline/main.nf b/subworkflows/nf-core/utils_nfcore_pipeline/main.nf new file mode 100644 index 00000000..14558c39 --- /dev/null +++ b/subworkflows/nf-core/utils_nfcore_pipeline/main.nf @@ -0,0 +1,446 @@ +// +// Subworkflow with utility functions specific to the nf-core pipeline template +// + +import org.yaml.snakeyaml.Yaml +import nextflow.extension.FilesEx + +/* +======================================================================================== + SUBWORKFLOW DEFINITION +======================================================================================== +*/ + +workflow UTILS_NFCORE_PIPELINE { + + take: + nextflow_cli_args + + main: + valid_config = checkConfigProvided() + checkProfileProvided(nextflow_cli_args) + + emit: + valid_config +} + +/* +======================================================================================== + FUNCTIONS +======================================================================================== +*/ + +// +// Warn if a -profile or Nextflow config has not been provided to run the pipeline +// +def checkConfigProvided() { + valid_config = true + if (workflow.profile == 'standard' && workflow.configFiles.size() <= 1) { + log.warn "[$workflow.manifest.name] You are attempting to run the pipeline without any custom configuration!\n\n" + + "This will be dependent on your local compute environment but can be achieved via one or more of the following:\n" + + " (1) Using an existing pipeline profile e.g. `-profile docker` or `-profile singularity`\n" + + " (2) Using an existing nf-core/configs for your Institution e.g. `-profile crick` or `-profile uppmax`\n" + + " (3) Using your own local custom config e.g. `-c /path/to/your/custom.config`\n\n" + + "Please refer to the quick start section and usage docs for the pipeline.\n " + valid_config = false + } + return valid_config +} + +// +// Exit pipeline if --profile contains spaces +// +def checkProfileProvided(nextflow_cli_args) { + if (workflow.profile.endsWith(',')) { + error "The `-profile` option cannot end with a trailing comma, please remove it and re-run the pipeline!\n" + + "HINT: A common mistake is to provide multiple values separated by spaces e.g. `-profile test, docker`.\n" + } + if (nextflow_cli_args[0]) { + log.warn "nf-core pipelines do not accept positional arguments. The positional argument `${nextflow_cli_args[0]}` has been detected.\n" + + "HINT: A common mistake is to provide multiple values separated by spaces e.g. `-profile test, docker`.\n" + } +} + +// +// Citation string for pipeline +// +def workflowCitation() { + def temp_doi_ref = "" + String[] manifest_doi = workflow.manifest.doi.tokenize(",") + // Using a loop to handle multiple DOIs + // Removing `https://doi.org/` to handle pipelines using DOIs vs DOI resolvers + // Removing ` ` since the manifest.doi is a string and not a proper list + for (String doi_ref: manifest_doi) temp_doi_ref += " https://doi.org/${doi_ref.replace('https://doi.org/', '').replace(' ', '')}\n" + return "If you use ${workflow.manifest.name} for your analysis please cite:\n\n" + + "* The pipeline\n" + + temp_doi_ref + "\n" + + "* The nf-core framework\n" + + " https://doi.org/10.1038/s41587-020-0439-x\n\n" + + "* Software dependencies\n" + + " https://github.com/${workflow.manifest.name}/blob/master/CITATIONS.md" +} + +// +// Generate workflow version string +// +def getWorkflowVersion() { + String version_string = "" + if (workflow.manifest.version) { + def prefix_v = workflow.manifest.version[0] != 'v' ? 'v' : '' + version_string += "${prefix_v}${workflow.manifest.version}" + } + + if (workflow.commitId) { + def git_shortsha = workflow.commitId.substring(0, 7) + version_string += "-g${git_shortsha}" + } + + return version_string +} + +// +// Get software versions for pipeline +// +def processVersionsFromYAML(yaml_file) { + Yaml yaml = new Yaml() + versions = yaml.load(yaml_file).collectEntries { k, v -> [ k.tokenize(':')[-1], v ] } + return yaml.dumpAsMap(versions).trim() +} + +// +// Get workflow version for pipeline +// +def workflowVersionToYAML() { + return """ + Workflow: + $workflow.manifest.name: ${getWorkflowVersion()} + Nextflow: $workflow.nextflow.version + """.stripIndent().trim() +} + +// +// Get channel of software versions used in pipeline in YAML format +// +def softwareVersionsToYAML(ch_versions) { + return ch_versions + .unique() + .map { processVersionsFromYAML(it) } + .unique() + .mix(Channel.of(workflowVersionToYAML())) +} + +// +// Get workflow summary for MultiQC +// +def paramsSummaryMultiqc(summary_params) { + def summary_section = '' + for (group in summary_params.keySet()) { + def group_params = summary_params.get(group) // This gets the parameters of that particular group + if (group_params) { + summary_section += "

    $group

    \n" + summary_section += "
    \n" + for (param in group_params.keySet()) { + summary_section += "
    $param
    ${group_params.get(param) ?: 'N/A'}
    \n" + } + summary_section += "
    \n" + } + } + + String yaml_file_text = "id: '${workflow.manifest.name.replace('/','-')}-summary'\n" + yaml_file_text += "description: ' - this information is collected when the pipeline is started.'\n" + yaml_file_text += "section_name: '${workflow.manifest.name} Workflow Summary'\n" + yaml_file_text += "section_href: 'https://github.com/${workflow.manifest.name}'\n" + yaml_file_text += "plot_type: 'html'\n" + yaml_file_text += "data: |\n" + yaml_file_text += "${summary_section}" + + return yaml_file_text +} + +// +// nf-core logo +// +def nfCoreLogo(monochrome_logs=true) { + Map colors = logColours(monochrome_logs) + String.format( + """\n + ${dashedLine(monochrome_logs)} + ${colors.green},--.${colors.black}/${colors.green},-.${colors.reset} + ${colors.blue} ___ __ __ __ ___ ${colors.green}/,-._.--~\'${colors.reset} + ${colors.blue} |\\ | |__ __ / ` / \\ |__) |__ ${colors.yellow}} {${colors.reset} + ${colors.blue} | \\| | \\__, \\__/ | \\ |___ ${colors.green}\\`-._,-`-,${colors.reset} + ${colors.green}`._,._,\'${colors.reset} + ${colors.purple} ${workflow.manifest.name} ${getWorkflowVersion()}${colors.reset} + ${dashedLine(monochrome_logs)} + """.stripIndent() + ) +} + +// +// Return dashed line +// +def dashedLine(monochrome_logs=true) { + Map colors = logColours(monochrome_logs) + return "-${colors.dim}----------------------------------------------------${colors.reset}-" +} + +// +// ANSII colours used for terminal logging +// +def logColours(monochrome_logs=true) { + Map colorcodes = [:] + + // Reset / Meta + colorcodes['reset'] = monochrome_logs ? '' : "\033[0m" + colorcodes['bold'] = monochrome_logs ? '' : "\033[1m" + colorcodes['dim'] = monochrome_logs ? '' : "\033[2m" + colorcodes['underlined'] = monochrome_logs ? '' : "\033[4m" + colorcodes['blink'] = monochrome_logs ? '' : "\033[5m" + colorcodes['reverse'] = monochrome_logs ? '' : "\033[7m" + colorcodes['hidden'] = monochrome_logs ? '' : "\033[8m" + + // Regular Colors + colorcodes['black'] = monochrome_logs ? '' : "\033[0;30m" + colorcodes['red'] = monochrome_logs ? '' : "\033[0;31m" + colorcodes['green'] = monochrome_logs ? '' : "\033[0;32m" + colorcodes['yellow'] = monochrome_logs ? '' : "\033[0;33m" + colorcodes['blue'] = monochrome_logs ? '' : "\033[0;34m" + colorcodes['purple'] = monochrome_logs ? '' : "\033[0;35m" + colorcodes['cyan'] = monochrome_logs ? '' : "\033[0;36m" + colorcodes['white'] = monochrome_logs ? '' : "\033[0;37m" + + // Bold + colorcodes['bblack'] = monochrome_logs ? '' : "\033[1;30m" + colorcodes['bred'] = monochrome_logs ? '' : "\033[1;31m" + colorcodes['bgreen'] = monochrome_logs ? '' : "\033[1;32m" + colorcodes['byellow'] = monochrome_logs ? '' : "\033[1;33m" + colorcodes['bblue'] = monochrome_logs ? '' : "\033[1;34m" + colorcodes['bpurple'] = monochrome_logs ? '' : "\033[1;35m" + colorcodes['bcyan'] = monochrome_logs ? '' : "\033[1;36m" + colorcodes['bwhite'] = monochrome_logs ? '' : "\033[1;37m" + + // Underline + colorcodes['ublack'] = monochrome_logs ? '' : "\033[4;30m" + colorcodes['ured'] = monochrome_logs ? '' : "\033[4;31m" + colorcodes['ugreen'] = monochrome_logs ? '' : "\033[4;32m" + colorcodes['uyellow'] = monochrome_logs ? '' : "\033[4;33m" + colorcodes['ublue'] = monochrome_logs ? '' : "\033[4;34m" + colorcodes['upurple'] = monochrome_logs ? '' : "\033[4;35m" + colorcodes['ucyan'] = monochrome_logs ? '' : "\033[4;36m" + colorcodes['uwhite'] = monochrome_logs ? '' : "\033[4;37m" + + // High Intensity + colorcodes['iblack'] = monochrome_logs ? '' : "\033[0;90m" + colorcodes['ired'] = monochrome_logs ? '' : "\033[0;91m" + colorcodes['igreen'] = monochrome_logs ? '' : "\033[0;92m" + colorcodes['iyellow'] = monochrome_logs ? '' : "\033[0;93m" + colorcodes['iblue'] = monochrome_logs ? '' : "\033[0;94m" + colorcodes['ipurple'] = monochrome_logs ? '' : "\033[0;95m" + colorcodes['icyan'] = monochrome_logs ? '' : "\033[0;96m" + colorcodes['iwhite'] = monochrome_logs ? '' : "\033[0;97m" + + // Bold High Intensity + colorcodes['biblack'] = monochrome_logs ? '' : "\033[1;90m" + colorcodes['bired'] = monochrome_logs ? '' : "\033[1;91m" + colorcodes['bigreen'] = monochrome_logs ? '' : "\033[1;92m" + colorcodes['biyellow'] = monochrome_logs ? '' : "\033[1;93m" + colorcodes['biblue'] = monochrome_logs ? '' : "\033[1;94m" + colorcodes['bipurple'] = monochrome_logs ? '' : "\033[1;95m" + colorcodes['bicyan'] = monochrome_logs ? '' : "\033[1;96m" + colorcodes['biwhite'] = monochrome_logs ? '' : "\033[1;97m" + + return colorcodes +} + +// +// Attach the multiqc report to email +// +def attachMultiqcReport(multiqc_report) { + def mqc_report = null + try { + if (workflow.success) { + mqc_report = multiqc_report.getVal() + if (mqc_report.getClass() == ArrayList && mqc_report.size() >= 1) { + if (mqc_report.size() > 1) { + log.warn "[$workflow.manifest.name] Found multiple reports from process 'MULTIQC', will use only one" + } + mqc_report = mqc_report[0] + } + } + } catch (all) { + if (multiqc_report) { + log.warn "[$workflow.manifest.name] Could not attach MultiQC report to summary email" + } + } + return mqc_report +} + +// +// Construct and send completion email +// +def completionEmail(summary_params, email, email_on_fail, plaintext_email, outdir, monochrome_logs=true, multiqc_report=null) { + + // Set up the e-mail variables + def subject = "[$workflow.manifest.name] Successful: $workflow.runName" + if (!workflow.success) { + subject = "[$workflow.manifest.name] FAILED: $workflow.runName" + } + + def summary = [:] + for (group in summary_params.keySet()) { + summary << summary_params[group] + } + + def misc_fields = [:] + misc_fields['Date Started'] = workflow.start + misc_fields['Date Completed'] = workflow.complete + misc_fields['Pipeline script file path'] = workflow.scriptFile + misc_fields['Pipeline script hash ID'] = workflow.scriptId + if (workflow.repository) misc_fields['Pipeline repository Git URL'] = workflow.repository + if (workflow.commitId) misc_fields['Pipeline repository Git Commit'] = workflow.commitId + if (workflow.revision) misc_fields['Pipeline Git branch/tag'] = workflow.revision + misc_fields['Nextflow Version'] = workflow.nextflow.version + misc_fields['Nextflow Build'] = workflow.nextflow.build + misc_fields['Nextflow Compile Timestamp'] = workflow.nextflow.timestamp + + def email_fields = [:] + email_fields['version'] = getWorkflowVersion() + email_fields['runName'] = workflow.runName + email_fields['success'] = workflow.success + email_fields['dateComplete'] = workflow.complete + email_fields['duration'] = workflow.duration + email_fields['exitStatus'] = workflow.exitStatus + email_fields['errorMessage'] = (workflow.errorMessage ?: 'None') + email_fields['errorReport'] = (workflow.errorReport ?: 'None') + email_fields['commandLine'] = workflow.commandLine + email_fields['projectDir'] = workflow.projectDir + email_fields['summary'] = summary << misc_fields + + // On success try attach the multiqc report + def mqc_report = attachMultiqcReport(multiqc_report) + + // Check if we are only sending emails on failure + def email_address = email + if (!email && email_on_fail && !workflow.success) { + email_address = email_on_fail + } + + // Render the TXT template + def engine = new groovy.text.GStringTemplateEngine() + def tf = new File("${workflow.projectDir}/assets/email_template.txt") + def txt_template = engine.createTemplate(tf).make(email_fields) + def email_txt = txt_template.toString() + + // Render the HTML template + def hf = new File("${workflow.projectDir}/assets/email_template.html") + def html_template = engine.createTemplate(hf).make(email_fields) + def email_html = html_template.toString() + + // Render the sendmail template + def max_multiqc_email_size = (params.containsKey('max_multiqc_email_size') ? params.max_multiqc_email_size : 0) as nextflow.util.MemoryUnit + def smail_fields = [ email: email_address, subject: subject, email_txt: email_txt, email_html: email_html, projectDir: "${workflow.projectDir}", mqcFile: mqc_report, mqcMaxSize: max_multiqc_email_size.toBytes() ] + def sf = new File("${workflow.projectDir}/assets/sendmail_template.txt") + def sendmail_template = engine.createTemplate(sf).make(smail_fields) + def sendmail_html = sendmail_template.toString() + + // Send the HTML e-mail + Map colors = logColours(monochrome_logs) + if (email_address) { + try { + if (plaintext_email) { throw GroovyException('Send plaintext e-mail, not HTML') } + // Try to send HTML e-mail using sendmail + def sendmail_tf = new File(workflow.launchDir.toString(), ".sendmail_tmp.html") + sendmail_tf.withWriter { w -> w << sendmail_html } + [ 'sendmail', '-t' ].execute() << sendmail_html + log.info "-${colors.purple}[$workflow.manifest.name]${colors.green} Sent summary e-mail to $email_address (sendmail)-" + } catch (all) { + // Catch failures and try with plaintext + def mail_cmd = [ 'mail', '-s', subject, '--content-type=text/html', email_address ] + mail_cmd.execute() << email_html + log.info "-${colors.purple}[$workflow.manifest.name]${colors.green} Sent summary e-mail to $email_address (mail)-" + } + } + + // Write summary e-mail HTML to a file + def output_hf = new File(workflow.launchDir.toString(), ".pipeline_report.html") + output_hf.withWriter { w -> w << email_html } + FilesEx.copyTo(output_hf.toPath(), "${outdir}/pipeline_info/pipeline_report.html"); + output_hf.delete() + + // Write summary e-mail TXT to a file + def output_tf = new File(workflow.launchDir.toString(), ".pipeline_report.txt") + output_tf.withWriter { w -> w << email_txt } + FilesEx.copyTo(output_tf.toPath(), "${outdir}/pipeline_info/pipeline_report.txt"); + output_tf.delete() +} + +// +// Print pipeline summary on completion +// +def completionSummary(monochrome_logs=true) { + Map colors = logColours(monochrome_logs) + if (workflow.success) { + if (workflow.stats.ignoredCount == 0) { + log.info "-${colors.purple}[$workflow.manifest.name]${colors.green} Pipeline completed successfully${colors.reset}-" + } else { + log.info "-${colors.purple}[$workflow.manifest.name]${colors.yellow} Pipeline completed successfully, but with errored process(es) ${colors.reset}-" + } + } else { + log.info "-${colors.purple}[$workflow.manifest.name]${colors.red} Pipeline completed with errors${colors.reset}-" + } +} + +// +// Construct and send a notification to a web server as JSON e.g. Microsoft Teams and Slack +// +def imNotification(summary_params, hook_url) { + def summary = [:] + for (group in summary_params.keySet()) { + summary << summary_params[group] + } + + def misc_fields = [:] + misc_fields['start'] = workflow.start + misc_fields['complete'] = workflow.complete + misc_fields['scriptfile'] = workflow.scriptFile + misc_fields['scriptid'] = workflow.scriptId + if (workflow.repository) misc_fields['repository'] = workflow.repository + if (workflow.commitId) misc_fields['commitid'] = workflow.commitId + if (workflow.revision) misc_fields['revision'] = workflow.revision + misc_fields['nxf_version'] = workflow.nextflow.version + misc_fields['nxf_build'] = workflow.nextflow.build + misc_fields['nxf_timestamp'] = workflow.nextflow.timestamp + + def msg_fields = [:] + msg_fields['version'] = getWorkflowVersion() + msg_fields['runName'] = workflow.runName + msg_fields['success'] = workflow.success + msg_fields['dateComplete'] = workflow.complete + msg_fields['duration'] = workflow.duration + msg_fields['exitStatus'] = workflow.exitStatus + msg_fields['errorMessage'] = (workflow.errorMessage ?: 'None') + msg_fields['errorReport'] = (workflow.errorReport ?: 'None') + msg_fields['commandLine'] = workflow.commandLine.replaceFirst(/ +--hook_url +[^ ]+/, "") + msg_fields['projectDir'] = workflow.projectDir + msg_fields['summary'] = summary << misc_fields + + // Render the JSON template + def engine = new groovy.text.GStringTemplateEngine() + // Different JSON depending on the service provider + // Defaults to "Adaptive Cards" (https://adaptivecards.io), except Slack which has its own format + def json_path = hook_url.contains("hooks.slack.com") ? "slackreport.json" : "adaptivecard.json" + def hf = new File("${workflow.projectDir}/assets/${json_path}") + def json_template = engine.createTemplate(hf).make(msg_fields) + def json_message = json_template.toString() + + // POST + def post = new URL(hook_url).openConnection(); + post.setRequestMethod("POST") + post.setDoOutput(true) + post.setRequestProperty("Content-Type", "application/json") + post.getOutputStream().write(json_message.getBytes("UTF-8")); + def postRC = post.getResponseCode(); + if (! postRC.equals(200)) { + log.warn(post.getErrorStream().getText()); + } +} diff --git a/subworkflows/nf-core/utils_nfcore_pipeline/meta.yml b/subworkflows/nf-core/utils_nfcore_pipeline/meta.yml new file mode 100644 index 00000000..d08d2434 --- /dev/null +++ b/subworkflows/nf-core/utils_nfcore_pipeline/meta.yml @@ -0,0 +1,24 @@ +# yaml-language-server: $schema=https://raw.githubusercontent.com/nf-core/modules/master/subworkflows/yaml-schema.json +name: "UTILS_NFCORE_PIPELINE" +description: Subworkflow with utility functions specific to the nf-core pipeline template +keywords: + - utility + - pipeline + - initialise + - version +components: [] +input: + - nextflow_cli_args: + type: list + description: | + Nextflow CLI positional arguments +output: + - success: + type: boolean + description: | + Dummy output to indicate success +authors: + - "@adamrtalbot" +maintainers: + - "@adamrtalbot" + - "@maxulysse" diff --git a/subworkflows/nf-core/utils_nfcore_pipeline/tests/main.function.nf.test b/subworkflows/nf-core/utils_nfcore_pipeline/tests/main.function.nf.test new file mode 100644 index 00000000..1dc317f8 --- /dev/null +++ b/subworkflows/nf-core/utils_nfcore_pipeline/tests/main.function.nf.test @@ -0,0 +1,134 @@ + +nextflow_function { + + name "Test Functions" + script "../main.nf" + config "subworkflows/nf-core/utils_nfcore_pipeline/tests/nextflow.config" + tag "subworkflows" + tag "subworkflows_nfcore" + tag "utils_nfcore_pipeline" + tag "subworkflows/utils_nfcore_pipeline" + + test("Test Function checkConfigProvided") { + + function "checkConfigProvided" + + then { + assertAll( + { assert function.success }, + { assert snapshot(function.result).match() } + ) + } + } + + test("Test Function checkProfileProvided") { + + function "checkProfileProvided" + + when { + function { + """ + input[0] = [] + """ + } + } + + then { + assertAll( + { assert function.success }, + { assert snapshot(function.result).match() } + ) + } + } + + test("Test Function workflowCitation") { + + function "workflowCitation" + + then { + assertAll( + { assert function.success }, + { assert snapshot(function.result).match() } + ) + } + } + + test("Test Function nfCoreLogo") { + + function "nfCoreLogo" + + when { + function { + """ + input[0] = false + """ + } + } + + then { + assertAll( + { assert function.success }, + { assert snapshot(function.result).match() } + ) + } + } + + test("Test Function dashedLine") { + + function "dashedLine" + + when { + function { + """ + input[0] = false + """ + } + } + + then { + assertAll( + { assert function.success }, + { assert snapshot(function.result).match() } + ) + } + } + + test("Test Function without logColours") { + + function "logColours" + + when { + function { + """ + input[0] = true + """ + } + } + + then { + assertAll( + { assert function.success }, + { assert snapshot(function.result).match() } + ) + } + } + + test("Test Function with logColours") { + function "logColours" + + when { + function { + """ + input[0] = false + """ + } + } + + then { + assertAll( + { assert function.success }, + { assert snapshot(function.result).match() } + ) + } + } +} diff --git a/subworkflows/nf-core/utils_nfcore_pipeline/tests/main.function.nf.test.snap b/subworkflows/nf-core/utils_nfcore_pipeline/tests/main.function.nf.test.snap new file mode 100644 index 00000000..1037232c --- /dev/null +++ b/subworkflows/nf-core/utils_nfcore_pipeline/tests/main.function.nf.test.snap @@ -0,0 +1,166 @@ +{ + "Test Function checkProfileProvided": { + "content": null, + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.10.1" + }, + "timestamp": "2024-02-28T12:03:03.360873" + }, + "Test Function checkConfigProvided": { + "content": [ + true + ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.10.1" + }, + "timestamp": "2024-02-28T12:02:59.729647" + }, + "Test Function nfCoreLogo": { + "content": [ + "\n\n-\u001b[2m----------------------------------------------------\u001b[0m-\n \u001b[0;32m,--.\u001b[0;30m/\u001b[0;32m,-.\u001b[0m\n\u001b[0;34m ___ __ __ __ ___ \u001b[0;32m/,-._.--~'\u001b[0m\n\u001b[0;34m |\\ | |__ __ / ` / \\ |__) |__ \u001b[0;33m} {\u001b[0m\n\u001b[0;34m | \\| | \\__, \\__/ | \\ |___ \u001b[0;32m\\`-._,-`-,\u001b[0m\n \u001b[0;32m`._,._,'\u001b[0m\n\u001b[0;35m nextflow_workflow v9.9.9\u001b[0m\n-\u001b[2m----------------------------------------------------\u001b[0m-\n" + ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.10.1" + }, + "timestamp": "2024-02-28T12:03:10.562934" + }, + "Test Function workflowCitation": { + "content": [ + "If you use nextflow_workflow for your analysis please cite:\n\n* The pipeline\n https://doi.org/10.5281/zenodo.5070524\n\n* The nf-core framework\n https://doi.org/10.1038/s41587-020-0439-x\n\n* Software dependencies\n https://github.com/nextflow_workflow/blob/master/CITATIONS.md" + ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.10.1" + }, + "timestamp": "2024-02-28T12:03:07.019761" + }, + "Test Function without logColours": { + "content": [ + { + "reset": "", + "bold": "", + "dim": "", + "underlined": "", + "blink": "", + "reverse": "", + "hidden": "", + "black": "", + "red": "", + "green": "", + "yellow": "", + "blue": "", + "purple": "", + "cyan": "", + "white": "", + "bblack": "", + "bred": "", + "bgreen": "", + "byellow": "", + "bblue": "", + "bpurple": "", + "bcyan": "", + "bwhite": "", + "ublack": "", + "ured": "", + "ugreen": "", + "uyellow": "", + "ublue": "", + "upurple": "", + "ucyan": "", + "uwhite": "", + "iblack": "", + "ired": "", + "igreen": "", + "iyellow": "", + "iblue": "", + "ipurple": "", + "icyan": "", + "iwhite": "", + "biblack": "", + "bired": "", + "bigreen": "", + "biyellow": "", + "biblue": "", + "bipurple": "", + "bicyan": "", + "biwhite": "" + } + ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.10.1" + }, + "timestamp": "2024-02-28T12:03:17.969323" + }, + "Test Function dashedLine": { + "content": [ + "-\u001b[2m----------------------------------------------------\u001b[0m-" + ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.10.1" + }, + "timestamp": "2024-02-28T12:03:14.366181" + }, + "Test Function with logColours": { + "content": [ + { + "reset": "\u001b[0m", + "bold": "\u001b[1m", + "dim": "\u001b[2m", + "underlined": "\u001b[4m", + "blink": "\u001b[5m", + "reverse": "\u001b[7m", + "hidden": "\u001b[8m", + "black": "\u001b[0;30m", + "red": "\u001b[0;31m", + "green": "\u001b[0;32m", + "yellow": "\u001b[0;33m", + "blue": "\u001b[0;34m", + "purple": "\u001b[0;35m", + "cyan": "\u001b[0;36m", + "white": "\u001b[0;37m", + "bblack": "\u001b[1;30m", + "bred": "\u001b[1;31m", + "bgreen": "\u001b[1;32m", + "byellow": "\u001b[1;33m", + "bblue": "\u001b[1;34m", + "bpurple": "\u001b[1;35m", + "bcyan": "\u001b[1;36m", + "bwhite": "\u001b[1;37m", + "ublack": "\u001b[4;30m", + "ured": "\u001b[4;31m", + "ugreen": "\u001b[4;32m", + "uyellow": "\u001b[4;33m", + "ublue": "\u001b[4;34m", + "upurple": "\u001b[4;35m", + "ucyan": "\u001b[4;36m", + "uwhite": "\u001b[4;37m", + "iblack": "\u001b[0;90m", + "ired": "\u001b[0;91m", + "igreen": "\u001b[0;92m", + "iyellow": "\u001b[0;93m", + "iblue": "\u001b[0;94m", + "ipurple": "\u001b[0;95m", + "icyan": "\u001b[0;96m", + "iwhite": "\u001b[0;97m", + "biblack": "\u001b[1;90m", + "bired": "\u001b[1;91m", + "bigreen": "\u001b[1;92m", + "biyellow": "\u001b[1;93m", + "biblue": "\u001b[1;94m", + "bipurple": "\u001b[1;95m", + "bicyan": "\u001b[1;96m", + "biwhite": "\u001b[1;97m" + } + ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.10.1" + }, + "timestamp": "2024-02-28T12:03:21.714424" + } +} \ No newline at end of file diff --git a/subworkflows/nf-core/utils_nfcore_pipeline/tests/main.workflow.nf.test b/subworkflows/nf-core/utils_nfcore_pipeline/tests/main.workflow.nf.test new file mode 100644 index 00000000..8940d32d --- /dev/null +++ b/subworkflows/nf-core/utils_nfcore_pipeline/tests/main.workflow.nf.test @@ -0,0 +1,29 @@ +nextflow_workflow { + + name "Test Workflow UTILS_NFCORE_PIPELINE" + script "../main.nf" + config "subworkflows/nf-core/utils_nfcore_pipeline/tests/nextflow.config" + workflow "UTILS_NFCORE_PIPELINE" + tag "subworkflows" + tag "subworkflows_nfcore" + tag "utils_nfcore_pipeline" + tag "subworkflows/utils_nfcore_pipeline" + + test("Should run without failures") { + + when { + workflow { + """ + input[0] = [] + """ + } + } + + then { + assertAll( + { assert workflow.success }, + { assert snapshot(workflow.out).match() } + ) + } + } +} diff --git a/subworkflows/nf-core/utils_nfcore_pipeline/tests/main.workflow.nf.test.snap b/subworkflows/nf-core/utils_nfcore_pipeline/tests/main.workflow.nf.test.snap new file mode 100644 index 00000000..859d1030 --- /dev/null +++ b/subworkflows/nf-core/utils_nfcore_pipeline/tests/main.workflow.nf.test.snap @@ -0,0 +1,19 @@ +{ + "Should run without failures": { + "content": [ + { + "0": [ + true + ], + "valid_config": [ + true + ] + } + ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.10.1" + }, + "timestamp": "2024-02-28T12:03:25.726491" + } +} \ No newline at end of file diff --git a/subworkflows/nf-core/utils_nfcore_pipeline/tests/nextflow.config b/subworkflows/nf-core/utils_nfcore_pipeline/tests/nextflow.config new file mode 100644 index 00000000..d0a926bf --- /dev/null +++ b/subworkflows/nf-core/utils_nfcore_pipeline/tests/nextflow.config @@ -0,0 +1,9 @@ +manifest { + name = 'nextflow_workflow' + author = """nf-core""" + homePage = 'https://127.0.0.1' + description = """Dummy pipeline""" + nextflowVersion = '!>=23.04.0' + version = '9.9.9' + doi = 'https://doi.org/10.5281/zenodo.5070524' +} diff --git a/subworkflows/nf-core/utils_nfcore_pipeline/tests/tags.yml b/subworkflows/nf-core/utils_nfcore_pipeline/tests/tags.yml new file mode 100644 index 00000000..ac8523c9 --- /dev/null +++ b/subworkflows/nf-core/utils_nfcore_pipeline/tests/tags.yml @@ -0,0 +1,2 @@ +subworkflows/utils_nfcore_pipeline: + - subworkflows/nf-core/utils_nfcore_pipeline/** diff --git a/subworkflows/nf-core/utils_nfvalidation_plugin/main.nf b/subworkflows/nf-core/utils_nfvalidation_plugin/main.nf new file mode 100644 index 00000000..2585b65d --- /dev/null +++ b/subworkflows/nf-core/utils_nfvalidation_plugin/main.nf @@ -0,0 +1,62 @@ +// +// Subworkflow that uses the nf-validation plugin to render help text and parameter summary +// + +/* +======================================================================================== + IMPORT NF-VALIDATION PLUGIN +======================================================================================== +*/ + +include { paramsHelp } from 'plugin/nf-validation' +include { paramsSummaryLog } from 'plugin/nf-validation' +include { validateParameters } from 'plugin/nf-validation' + +/* +======================================================================================== + SUBWORKFLOW DEFINITION +======================================================================================== +*/ + +workflow UTILS_NFVALIDATION_PLUGIN { + + take: + print_help // boolean: print help + workflow_command // string: default commmand used to run pipeline + pre_help_text // string: string to be printed before help text and summary log + post_help_text // string: string to be printed after help text and summary log + validate_params // boolean: validate parameters + schema_filename // path: JSON schema file, null to use default value + + main: + + log.debug "Using schema file: ${schema_filename}" + + // Default values for strings + pre_help_text = pre_help_text ?: '' + post_help_text = post_help_text ?: '' + workflow_command = workflow_command ?: '' + + // + // Print help message if needed + // + if (print_help) { + log.info pre_help_text + paramsHelp(workflow_command, parameters_schema: schema_filename) + post_help_text + System.exit(0) + } + + // + // Print parameter summary to stdout + // + log.info pre_help_text + paramsSummaryLog(workflow, parameters_schema: schema_filename) + post_help_text + + // + // Validate parameters relative to the parameter JSON schema + // + if (validate_params){ + validateParameters(parameters_schema: schema_filename) + } + + emit: + dummy_emit = true +} diff --git a/subworkflows/nf-core/utils_nfvalidation_plugin/meta.yml b/subworkflows/nf-core/utils_nfvalidation_plugin/meta.yml new file mode 100644 index 00000000..3d4a6b04 --- /dev/null +++ b/subworkflows/nf-core/utils_nfvalidation_plugin/meta.yml @@ -0,0 +1,44 @@ +# yaml-language-server: $schema=https://raw.githubusercontent.com/nf-core/modules/master/subworkflows/yaml-schema.json +name: "UTILS_NFVALIDATION_PLUGIN" +description: Use nf-validation to initiate and validate a pipeline +keywords: + - utility + - pipeline + - initialise + - validation +components: [] +input: + - print_help: + type: boolean + description: | + Print help message and exit + - workflow_command: + type: string + description: | + The command to run the workflow e.g. "nextflow run main.nf" + - pre_help_text: + type: string + description: | + Text to print before the help message + - post_help_text: + type: string + description: | + Text to print after the help message + - validate_params: + type: boolean + description: | + Validate the parameters and error if invalid. + - schema_filename: + type: string + description: | + The filename of the schema to validate against. +output: + - dummy_emit: + type: boolean + description: | + Dummy emit to make nf-core subworkflows lint happy +authors: + - "@adamrtalbot" +maintainers: + - "@adamrtalbot" + - "@maxulysse" diff --git a/subworkflows/nf-core/utils_nfvalidation_plugin/tests/main.nf.test b/subworkflows/nf-core/utils_nfvalidation_plugin/tests/main.nf.test new file mode 100644 index 00000000..5784a33f --- /dev/null +++ b/subworkflows/nf-core/utils_nfvalidation_plugin/tests/main.nf.test @@ -0,0 +1,200 @@ +nextflow_workflow { + + name "Test Workflow UTILS_NFVALIDATION_PLUGIN" + script "../main.nf" + workflow "UTILS_NFVALIDATION_PLUGIN" + tag "subworkflows" + tag "subworkflows_nfcore" + tag "plugin/nf-validation" + tag "'plugin/nf-validation'" + tag "utils_nfvalidation_plugin" + tag "subworkflows/utils_nfvalidation_plugin" + + test("Should run nothing") { + + when { + + params { + monochrome_logs = true + test_data = '' + } + + workflow { + """ + help = false + workflow_command = null + pre_help_text = null + post_help_text = null + validate_params = false + schema_filename = "$moduleTestDir/nextflow_schema.json" + + input[0] = help + input[1] = workflow_command + input[2] = pre_help_text + input[3] = post_help_text + input[4] = validate_params + input[5] = schema_filename + """ + } + } + + then { + assertAll( + { assert workflow.success } + ) + } + } + + test("Should run help") { + + + when { + + params { + monochrome_logs = true + test_data = '' + } + workflow { + """ + help = true + workflow_command = null + pre_help_text = null + post_help_text = null + validate_params = false + schema_filename = "$moduleTestDir/nextflow_schema.json" + + input[0] = help + input[1] = workflow_command + input[2] = pre_help_text + input[3] = post_help_text + input[4] = validate_params + input[5] = schema_filename + """ + } + } + + then { + assertAll( + { assert workflow.success }, + { assert workflow.exitStatus == 0 }, + { assert workflow.stdout.any { it.contains('Input/output options') } }, + { assert workflow.stdout.any { it.contains('--outdir') } } + ) + } + } + + test("Should run help with command") { + + when { + + params { + monochrome_logs = true + test_data = '' + } + workflow { + """ + help = true + workflow_command = "nextflow run noorg/doesntexist" + pre_help_text = null + post_help_text = null + validate_params = false + schema_filename = "$moduleTestDir/nextflow_schema.json" + + input[0] = help + input[1] = workflow_command + input[2] = pre_help_text + input[3] = post_help_text + input[4] = validate_params + input[5] = schema_filename + """ + } + } + + then { + assertAll( + { assert workflow.success }, + { assert workflow.exitStatus == 0 }, + { assert workflow.stdout.any { it.contains('nextflow run noorg/doesntexist') } }, + { assert workflow.stdout.any { it.contains('Input/output options') } }, + { assert workflow.stdout.any { it.contains('--outdir') } } + ) + } + } + + test("Should run help with extra text") { + + + when { + + params { + monochrome_logs = true + test_data = '' + } + workflow { + """ + help = true + workflow_command = "nextflow run noorg/doesntexist" + pre_help_text = "pre-help-text" + post_help_text = "post-help-text" + validate_params = false + schema_filename = "$moduleTestDir/nextflow_schema.json" + + input[0] = help + input[1] = workflow_command + input[2] = pre_help_text + input[3] = post_help_text + input[4] = validate_params + input[5] = schema_filename + """ + } + } + + then { + assertAll( + { assert workflow.success }, + { assert workflow.exitStatus == 0 }, + { assert workflow.stdout.any { it.contains('pre-help-text') } }, + { assert workflow.stdout.any { it.contains('nextflow run noorg/doesntexist') } }, + { assert workflow.stdout.any { it.contains('Input/output options') } }, + { assert workflow.stdout.any { it.contains('--outdir') } }, + { assert workflow.stdout.any { it.contains('post-help-text') } } + ) + } + } + + test("Should validate params") { + + when { + + params { + monochrome_logs = true + test_data = '' + outdir = 1 + } + workflow { + """ + help = false + workflow_command = null + pre_help_text = null + post_help_text = null + validate_params = true + schema_filename = "$moduleTestDir/nextflow_schema.json" + + input[0] = help + input[1] = workflow_command + input[2] = pre_help_text + input[3] = post_help_text + input[4] = validate_params + input[5] = schema_filename + """ + } + } + + then { + assertAll( + { assert workflow.failed }, + { assert workflow.stdout.any { it.contains('ERROR ~ ERROR: Validation of pipeline parameters failed!') } } + ) + } + } +} diff --git a/subworkflows/nf-core/utils_nfvalidation_plugin/tests/nextflow_schema.json b/subworkflows/nf-core/utils_nfvalidation_plugin/tests/nextflow_schema.json new file mode 100644 index 00000000..7626c1c9 --- /dev/null +++ b/subworkflows/nf-core/utils_nfvalidation_plugin/tests/nextflow_schema.json @@ -0,0 +1,96 @@ +{ + "$schema": "http://json-schema.org/draft-07/schema", + "$id": "https://raw.githubusercontent.com/./master/nextflow_schema.json", + "title": ". pipeline parameters", + "description": "", + "type": "object", + "definitions": { + "input_output_options": { + "title": "Input/output options", + "type": "object", + "fa_icon": "fas fa-terminal", + "description": "Define where the pipeline should find input data and save output data.", + "required": ["outdir"], + "properties": { + "validate_params": { + "type": "boolean", + "description": "Validate parameters?", + "default": true, + "hidden": true + }, + "outdir": { + "type": "string", + "format": "directory-path", + "description": "The output directory where the results will be saved. You have to use absolute paths to storage on Cloud infrastructure.", + "fa_icon": "fas fa-folder-open" + }, + "test_data_base": { + "type": "string", + "default": "https://raw.githubusercontent.com/nf-core/test-datasets/modules", + "description": "Base for test data directory", + "hidden": true + }, + "test_data": { + "type": "string", + "description": "Fake test data param", + "hidden": true + } + } + }, + "generic_options": { + "title": "Generic options", + "type": "object", + "fa_icon": "fas fa-file-import", + "description": "Less common options for the pipeline, typically set in a config file.", + "help_text": "These options are common to all nf-core pipelines and allow you to customise some of the core preferences for how the pipeline runs.\n\nTypically these options would be set in a Nextflow config file loaded for all pipeline runs, such as `~/.nextflow/config`.", + "properties": { + "help": { + "type": "boolean", + "description": "Display help text.", + "fa_icon": "fas fa-question-circle", + "hidden": true + }, + "version": { + "type": "boolean", + "description": "Display version and exit.", + "fa_icon": "fas fa-question-circle", + "hidden": true + }, + "logo": { + "type": "boolean", + "default": true, + "description": "Display nf-core logo in console output.", + "fa_icon": "fas fa-image", + "hidden": true + }, + "singularity_pull_docker_container": { + "type": "boolean", + "description": "Pull Singularity container from Docker?", + "hidden": true + }, + "publish_dir_mode": { + "type": "string", + "default": "copy", + "description": "Method used to save pipeline results to output directory.", + "help_text": "The Nextflow `publishDir` option specifies which intermediate files should be saved to the output directory. This option tells the pipeline what method should be used to move these files. See [Nextflow docs](https://www.nextflow.io/docs/latest/process.html#publishdir) for details.", + "fa_icon": "fas fa-copy", + "enum": ["symlink", "rellink", "link", "copy", "copyNoFollow", "move"], + "hidden": true + }, + "monochrome_logs": { + "type": "boolean", + "description": "Use monochrome_logs", + "hidden": true + } + } + } + }, + "allOf": [ + { + "$ref": "#/definitions/input_output_options" + }, + { + "$ref": "#/definitions/generic_options" + } + ] +} diff --git a/subworkflows/nf-core/utils_nfvalidation_plugin/tests/tags.yml b/subworkflows/nf-core/utils_nfvalidation_plugin/tests/tags.yml new file mode 100644 index 00000000..60b1cfff --- /dev/null +++ b/subworkflows/nf-core/utils_nfvalidation_plugin/tests/tags.yml @@ -0,0 +1,2 @@ +subworkflows/utils_nfvalidation_plugin: + - subworkflows/nf-core/utils_nfvalidation_plugin/** diff --git a/tower.yml b/tower.yml new file mode 100644 index 00000000..787aedfe --- /dev/null +++ b/tower.yml @@ -0,0 +1,5 @@ +reports: + multiqc_report.html: + display: "MultiQC HTML report" + samplesheet.csv: + display: "Auto-created samplesheet with collated metadata and FASTQ paths" diff --git a/workflows/rnadeseq.nf b/workflows/rnadeseq.nf new file mode 100644 index 00000000..b71412b7 --- /dev/null +++ b/workflows/rnadeseq.nf @@ -0,0 +1,98 @@ +/* +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ + IMPORT MODULES / SUBWORKFLOWS / FUNCTIONS +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ +*/ + +include { FASTQC } from '../modules/nf-core/fastqc/main' +include { MULTIQC } from '../modules/nf-core/multiqc/main' +include { paramsSummaryMap } from 'plugin/nf-validation' +include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' +include { softwareVersionsToYAML } from '../subworkflows/nf-core/utils_nfcore_pipeline' +include { methodsDescriptionText } from '../subworkflows/local/utils_nfcore_rnadeseq_pipeline' + +/* +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ + RUN MAIN WORKFLOW +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ +*/ + +workflow RNADESEQ { + + take: + ch_samplesheet // channel: samplesheet read in from --input + + main: + + ch_versions = Channel.empty() + ch_multiqc_files = Channel.empty() + + // + // MODULE: Run FastQC + // + FASTQC ( + ch_samplesheet + ) + ch_multiqc_files = ch_multiqc_files.mix(FASTQC.out.zip.collect{it[1]}) + ch_versions = ch_versions.mix(FASTQC.out.versions.first()) + + // + // Collate and save software versions + // + softwareVersionsToYAML(ch_versions) + .collectFile( + storeDir: "${params.outdir}/pipeline_info", + name: 'nf_core_pipeline_software_mqc_versions.yml', + sort: true, + newLine: true + ).set { ch_collated_versions } + + // + // MODULE: MultiQC + // + ch_multiqc_config = Channel.fromPath( + "$projectDir/assets/multiqc_config.yml", checkIfExists: true) + ch_multiqc_custom_config = params.multiqc_config ? + Channel.fromPath(params.multiqc_config, checkIfExists: true) : + Channel.empty() + ch_multiqc_logo = params.multiqc_logo ? + Channel.fromPath(params.multiqc_logo, checkIfExists: true) : + Channel.empty() + + summary_params = paramsSummaryMap( + workflow, parameters_schema: "nextflow_schema.json") + ch_workflow_summary = Channel.value(paramsSummaryMultiqc(summary_params)) + + ch_multiqc_custom_methods_description = params.multiqc_methods_description ? + file(params.multiqc_methods_description, checkIfExists: true) : + file("$projectDir/assets/methods_description_template.yml", checkIfExists: true) + ch_methods_description = Channel.value( + methodsDescriptionText(ch_multiqc_custom_methods_description)) + + ch_multiqc_files = ch_multiqc_files.mix( + ch_workflow_summary.collectFile(name: 'workflow_summary_mqc.yaml')) + ch_multiqc_files = ch_multiqc_files.mix(ch_collated_versions) + ch_multiqc_files = ch_multiqc_files.mix( + ch_methods_description.collectFile( + name: 'methods_description_mqc.yaml', + sort: true + ) + ) + + MULTIQC ( + ch_multiqc_files.collect(), + ch_multiqc_config.toList(), + ch_multiqc_custom_config.toList(), + ch_multiqc_logo.toList() + ) + + emit: + multiqc_report = MULTIQC.out.report.toList() // channel: /path/to/multiqc_report.html + versions = ch_versions // channel: [ path(versions.yml) ] +} + +/* +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ + THE END +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ +*/ From a804c98165c3ab382ae719c37c400202fc97fa1a Mon Sep 17 00:00:00 2001 From: WackerO Date: Thu, 16 May 2024 09:32:03 +0200 Subject: [PATCH 53/73] Further changes for new template, deleted some files, removed some old function calls --- .nf-core.yml | 24 +-- assets/RNAseq_report.Rmd | 4 +- assets/multiqc_config.yml | 2 +- bin/markdown_to_html.r | 51 ----- lib/NfcoreTemplate.groovy | 347 ------------------------------ lib/Utils.groovy | 47 ---- lib/WorkflowMain.groovy | 63 ------ lib/WorkflowRnadeseq.groovy | 118 ---------- lib/nfcore_external_java_deps.jar | Bin 2291171 -> 0 bytes main.nf | 25 +-- nextflow.config | 3 + nextflow_schema.json | 14 +- workflows/rnadeseq.nf | 31 +-- 13 files changed, 23 insertions(+), 706 deletions(-) delete mode 100755 bin/markdown_to_html.r delete mode 100755 lib/NfcoreTemplate.groovy delete mode 100755 lib/Utils.groovy delete mode 100755 lib/WorkflowMain.groovy delete mode 100755 lib/WorkflowRnadeseq.groovy delete mode 100644 lib/nfcore_external_java_deps.jar diff --git a/.nf-core.yml b/.nf-core.yml index f6fbb8af..f763727e 100644 --- a/.nf-core.yml +++ b/.nf-core.yml @@ -5,35 +5,19 @@ lint: files_unchanged: - .github/CONTRIBUTING.md - .github/ISSUE_TEMPLATE/bug_report.yml - - .github/ISSUE_TEMPLATE/config.yml - - .github/ISSUE_TEMPLATE/feature_request.yml - .github/PULL_REQUEST_TEMPLATE.md - .github/workflows/branch.yml - - .github/workflows/linting.yml - .gitignore - - assets/email_template.html - assets/email_template.txt - assets/sendmail_template.txt - docs/README.md - LICENSE - files_exist: - - .github/ISSUE_TEMPLATE/config.yml - - .github/workflows/awsfulltest.yml - - .github/workflows/awstest.yml - - assets/multiqc_config.yaml - - assets/nf-core-qbic-pipelines/rnadeseq_logo_light.png - - bin/markdown_to_html.r - - conf/test_full.config - - docs/images/nf-core-qbic-pipelines/rnadeseq_logo_dark.png - - docs/images/nf-core-qbic-pipelines/rnadeseq_logo_light.png - - lib/WorkflowQbic-pipelines/rnadeseq.groovy - nextflow_config: - - manifest.homePage - manifest.name - - params.input - - show_hidden_params + - manifest.homePage + - config_defaults: + - params.custom_config_base multiqc_config: - - report_comment # otherwise throws error: multiqc_config: 'assets/multiqc_config.yml' does not contain a matching 'report_comment'. │ + - report_comment # otherwise throws error: multiqc_config: 'assets/multiqc_config.yml' does not contain a matching 'report_comment'. diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index 3717035a..8d79f7f8 100644 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -2608,7 +2608,7 @@ as.character(params$revision), "` pipeline [^5], which was written using the nf-core template [@ewels2020nf]. For differential expression analysis, the read quantification data resulting from `", quant_tool, "` were processed with the R package `DESeq2 v", packageVersion("DESeq2"), -"` [@love2014differential]. The thresholds for differentially expressed genes were set to ", +"` [@love2014differential]. The thresholds for differentially expressed genes were set to ", pval_text, " for the adjusted p-value and ", logFC_text, " for the log2 Fold Change." )) @@ -2660,7 +2660,7 @@ The following lists all packages used for analysis, together with their versions ```{r versions} sessioninfo::session_info()%>% - details::details(summary = 'Session info/used packages') + details::details(summary = 'Session info/used packages') ``` --- diff --git a/assets/multiqc_config.yml b/assets/multiqc_config.yml index e9844102..439b820e 100644 --- a/assets/multiqc_config.yml +++ b/assets/multiqc_config.yml @@ -7,7 +7,7 @@ report_section_order: order: -1000 software_versions: order: -1001 - "nf-core-rnadeseq-summary": + "qbic-pipelines-rnadeseq-summary": order: -1002 export_plots: true diff --git a/bin/markdown_to_html.r b/bin/markdown_to_html.r deleted file mode 100755 index 212d793c..00000000 --- a/bin/markdown_to_html.r +++ /dev/null @@ -1,51 +0,0 @@ -#!/usr/bin/env Rscript - -# Command line argument processing -args = commandArgs(trailingOnly=TRUE) -if (length(args) < 2) { - stop("Usage: markdown_to_html.r ", call.=FALSE) -} -markdown_fn <- args[1] -output_fn <- args[2] - -# Load / install packages -if (!require("markdown")) { - install.packages("markdown", dependencies=TRUE, repos='http://cloud.r-project.org/') - library("markdown") -} - -base_css_fn <- getOption("markdown.HTML.stylesheet") -base_css <- readChar(base_css_fn, file.info(base_css_fn)$size) -custom_css <- paste(base_css, " -body { - padding: 3em; - margin-right: 350px; - max-width: 100%; -} -#toc { - position: fixed; - right: 20px; - width: 300px; - padding-top: 20px; - overflow: scroll; - height: calc(100% - 3em - 20px); -} -#toc_header { - font-size: 1.8em; - font-weight: bold; -} -#toc > ul { - padding-left: 0; - list-style-type: none; -} -#toc > ul ul { padding-left: 20px; } -#toc > ul > li > a { display: none; } -img { max-width: 800px; } -") - -markdownToHTML( - file = markdown_fn, - output = output_fn, - stylesheet = custom_css, - options = c('toc', 'base64_images', 'highlight_code') -) diff --git a/lib/NfcoreTemplate.groovy b/lib/NfcoreTemplate.groovy deleted file mode 100755 index c084c800..00000000 --- a/lib/NfcoreTemplate.groovy +++ /dev/null @@ -1,347 +0,0 @@ -// -// This file holds several functions used within the nf-core pipeline template. -// - -import org.yaml.snakeyaml.Yaml -import groovy.json.JsonOutput - -class NfcoreTemplate { - - // - // Check AWS Batch related parameters have been specified correctly - // - public static void awsBatch(workflow, params) { - if (workflow.profile.contains('awsbatch')) { - // Check params.awsqueue and params.awsregion have been set if running on AWSBatch - assert (params.awsqueue && params.awsregion) : "Specify correct --awsqueue and --awsregion parameters on AWSBatch!" - // Check outdir paths to be S3 buckets if running on AWSBatch - assert params.outdir.startsWith('s3:') : "Outdir not on S3 - specify S3 Bucket to run on AWSBatch!" - } - } - - // - // Warn if a -profile or Nextflow config has not been provided to run the pipeline - // - public static void checkConfigProvided(workflow, log) { - if (workflow.profile == 'standard' && workflow.configFiles.size() <= 1) { - log.warn "[$workflow.manifest.name] You are attempting to run the pipeline without any custom configuration!\n\n" + - "This will be dependent on your local compute environment but can be achieved via one or more of the following:\n" + - " (1) Using an existing pipeline profile e.g. `-profile docker` or `-profile singularity`\n" + - " (2) Using an existing nf-core/configs for your Institution e.g. `-profile crick` or `-profile uppmax`\n" + - " (3) Using your own local custom config e.g. `-c /path/to/your/custom.config`\n\n" + - "Please refer to the quick start section and usage docs for the pipeline.\n " - } - } - - // - // Generate version string - // - public static String version(workflow) { - String version_string = "" - - if (workflow.manifest.version) { - def prefix_v = workflow.manifest.version[0] != 'v' ? 'v' : '' - version_string += "${prefix_v}${workflow.manifest.version}" - } - - if (workflow.commitId) { - def git_shortsha = workflow.commitId.substring(0, 7) - version_string += "-g${git_shortsha}" - } - - return version_string - } - - // - // Construct and send completion email - // - public static void email(workflow, params, summary_params, projectDir, log, multiqc_report=[]) { - - // Set up the e-mail variables - def subject = "[$workflow.manifest.name] Successful: $workflow.runName" - if (!workflow.success) { - subject = "[$workflow.manifest.name] FAILED: $workflow.runName" - } - - def summary = [:] - for (group in summary_params.keySet()) { - summary << summary_params[group] - } - - def misc_fields = [:] - misc_fields['Date Started'] = workflow.start - misc_fields['Date Completed'] = workflow.complete - misc_fields['Pipeline script file path'] = workflow.scriptFile - misc_fields['Pipeline script hash ID'] = workflow.scriptId - if (workflow.repository) misc_fields['Pipeline repository Git URL'] = workflow.repository - if (workflow.commitId) misc_fields['Pipeline repository Git Commit'] = workflow.commitId - if (workflow.revision) misc_fields['Pipeline Git branch/tag'] = workflow.revision - misc_fields['Nextflow Version'] = workflow.nextflow.version - misc_fields['Nextflow Build'] = workflow.nextflow.build - misc_fields['Nextflow Compile Timestamp'] = workflow.nextflow.timestamp - - def email_fields = [:] - email_fields['version'] = NfcoreTemplate.version(workflow) - email_fields['runName'] = workflow.runName - email_fields['success'] = workflow.success - email_fields['dateComplete'] = workflow.complete - email_fields['duration'] = workflow.duration - email_fields['exitStatus'] = workflow.exitStatus - email_fields['errorMessage'] = (workflow.errorMessage ?: 'None') - email_fields['errorReport'] = (workflow.errorReport ?: 'None') - email_fields['commandLine'] = workflow.commandLine - email_fields['projectDir'] = workflow.projectDir - email_fields['summary'] = summary << misc_fields - - // On success try attach the multiqc report - def mqc_report = null - try { - if (workflow.success) { - mqc_report = multiqc_report.getVal() - if (mqc_report.getClass() == ArrayList && mqc_report.size() >= 1) { - if (mqc_report.size() > 1) { - log.warn "[$workflow.manifest.name] Found multiple reports from process 'MULTIQC', will use only one" - } - mqc_report = mqc_report[0] - } - } - } catch (all) { - if (multiqc_report) { - log.warn "[$workflow.manifest.name] Could not attach MultiQC report to summary email" - } - } - - // Check if we are only sending emails on failure - def email_address = params.email - if (!params.email && params.email_on_fail && !workflow.success) { - email_address = params.email_on_fail - } - - // Render the TXT template - def engine = new groovy.text.GStringTemplateEngine() - def tf = new File("$projectDir/assets/email_template.txt") - def txt_template = engine.createTemplate(tf).make(email_fields) - def email_txt = txt_template.toString() - - // Render the HTML template - def hf = new File("$projectDir/assets/email_template.html") - def html_template = engine.createTemplate(hf).make(email_fields) - def email_html = html_template.toString() - - // Render the sendmail template - def max_multiqc_email_size = (params.containsKey('max_multiqc_email_size') ? params.max_multiqc_email_size : 0) as nextflow.util.MemoryUnit - def smail_fields = [ email: email_address, subject: subject, email_txt: email_txt, email_html: email_html, projectDir: "$projectDir", mqcFile: mqc_report, mqcMaxSize: max_multiqc_email_size.toBytes() ] - def sf = new File("$projectDir/assets/sendmail_template.txt") - def sendmail_template = engine.createTemplate(sf).make(smail_fields) - def sendmail_html = sendmail_template.toString() - - // Send the HTML e-mail - Map colors = logColours(params.monochrome_logs) - if (email_address) { - try { - if (params.plaintext_email) { throw GroovyException('Send plaintext e-mail, not HTML') } - // Try to send HTML e-mail using sendmail - [ 'sendmail', '-t' ].execute() << sendmail_html - log.info "-${colors.purple}[$workflow.manifest.name]${colors.green} Sent summary e-mail to $email_address (sendmail)-" - } catch (all) { - // Catch failures and try with plaintext - def mail_cmd = [ 'mail', '-s', subject, '--content-type=text/html', email_address ] - if ( mqc_report.size() <= max_multiqc_email_size.toBytes() ) { - mail_cmd += [ '-A', mqc_report ] - } - mail_cmd.execute() << email_html - log.info "-${colors.purple}[$workflow.manifest.name]${colors.green} Sent summary e-mail to $email_address (mail)-" - } - } - - // Write summary e-mail HTML to a file - def output_d = new File("${params.outdir}/pipeline_info/") - if (!output_d.exists()) { - output_d.mkdirs() - } - def output_hf = new File(output_d, "pipeline_report.html") - output_hf.withWriter { w -> w << email_html } - def output_tf = new File(output_d, "pipeline_report.txt") - output_tf.withWriter { w -> w << email_txt } - } - - // - // Construct and send a notification to a web server as JSON - // e.g. Microsoft Teams and Slack - // - public static void IM_notification(workflow, params, summary_params, projectDir, log) { - def hook_url = params.hook_url - - def summary = [:] - for (group in summary_params.keySet()) { - summary << summary_params[group] - } - - def misc_fields = [:] - misc_fields['start'] = workflow.start - misc_fields['complete'] = workflow.complete - misc_fields['scriptfile'] = workflow.scriptFile - misc_fields['scriptid'] = workflow.scriptId - if (workflow.repository) misc_fields['repository'] = workflow.repository - if (workflow.commitId) misc_fields['commitid'] = workflow.commitId - if (workflow.revision) misc_fields['revision'] = workflow.revision - misc_fields['nxf_version'] = workflow.nextflow.version - misc_fields['nxf_build'] = workflow.nextflow.build - misc_fields['nxf_timestamp'] = workflow.nextflow.timestamp - - def msg_fields = [:] - msg_fields['version'] = NfcoreTemplate.version(workflow) - msg_fields['runName'] = workflow.runName - msg_fields['success'] = workflow.success - msg_fields['dateComplete'] = workflow.complete - msg_fields['duration'] = workflow.duration - msg_fields['exitStatus'] = workflow.exitStatus - msg_fields['errorMessage'] = (workflow.errorMessage ?: 'None') - msg_fields['errorReport'] = (workflow.errorReport ?: 'None') - msg_fields['commandLine'] = workflow.commandLine.replaceFirst(/ +--hook_url +[^ ]+/, "") - msg_fields['projectDir'] = workflow.projectDir - msg_fields['summary'] = summary << misc_fields - - // Render the JSON template - def engine = new groovy.text.GStringTemplateEngine() - // Different JSON depending on the service provider - // Defaults to "Adaptive Cards" (https://adaptivecards.io), except Slack which has its own format - def json_path = hook_url.contains("hooks.slack.com") ? "slackreport.json" : "adaptivecard.json" - def hf = new File("$projectDir/assets/${json_path}") - def json_template = engine.createTemplate(hf).make(msg_fields) - def json_message = json_template.toString() - - // POST - def post = new URL(hook_url).openConnection(); - post.setRequestMethod("POST") - post.setDoOutput(true) - post.setRequestProperty("Content-Type", "application/json") - post.getOutputStream().write(json_message.getBytes("UTF-8")); - def postRC = post.getResponseCode(); - if (! postRC.equals(200)) { - log.warn(post.getErrorStream().getText()); - } - } - - // - // Dump pipeline parameters in a json file - // - public static void dump_parameters(workflow, params) { - def output_d = new File("${params.outdir}/pipeline_info/") - if (!output_d.exists()) { - output_d.mkdirs() - } - - def timestamp = new java.util.Date().format( 'yyyy-MM-dd_HH-mm-ss') - def output_pf = new File(output_d, "params_${timestamp}.json") - def jsonStr = JsonOutput.toJson(params) - output_pf.text = JsonOutput.prettyPrint(jsonStr) - } - - // - // Print pipeline summary on completion - // - public static void summary(workflow, params, log) { - Map colors = logColours(params.monochrome_logs) - if (workflow.success) { - if (workflow.stats.ignoredCount == 0) { - log.info "-${colors.purple}[$workflow.manifest.name]${colors.green} Pipeline completed successfully${colors.reset}-" - } else { - log.info "-${colors.purple}[$workflow.manifest.name]${colors.yellow} Pipeline completed successfully, but with errored process(es) ${colors.reset}-" - } - } else { - log.info "-${colors.purple}[$workflow.manifest.name]${colors.red} Pipeline completed with errors${colors.reset}-" - } - } - - // - // ANSII Colours used for terminal logging - // - public static Map logColours(Boolean monochrome_logs) { - Map colorcodes = [:] - - // Reset / Meta - colorcodes['reset'] = monochrome_logs ? '' : "\033[0m" - colorcodes['bold'] = monochrome_logs ? '' : "\033[1m" - colorcodes['dim'] = monochrome_logs ? '' : "\033[2m" - colorcodes['underlined'] = monochrome_logs ? '' : "\033[4m" - colorcodes['blink'] = monochrome_logs ? '' : "\033[5m" - colorcodes['reverse'] = monochrome_logs ? '' : "\033[7m" - colorcodes['hidden'] = monochrome_logs ? '' : "\033[8m" - - // Regular Colors - colorcodes['black'] = monochrome_logs ? '' : "\033[0;30m" - colorcodes['red'] = monochrome_logs ? '' : "\033[0;31m" - colorcodes['green'] = monochrome_logs ? '' : "\033[0;32m" - colorcodes['yellow'] = monochrome_logs ? '' : "\033[0;33m" - colorcodes['blue'] = monochrome_logs ? '' : "\033[0;34m" - colorcodes['purple'] = monochrome_logs ? '' : "\033[0;35m" - colorcodes['cyan'] = monochrome_logs ? '' : "\033[0;36m" - colorcodes['white'] = monochrome_logs ? '' : "\033[0;37m" - - // Bold - colorcodes['bblack'] = monochrome_logs ? '' : "\033[1;30m" - colorcodes['bred'] = monochrome_logs ? '' : "\033[1;31m" - colorcodes['bgreen'] = monochrome_logs ? '' : "\033[1;32m" - colorcodes['byellow'] = monochrome_logs ? '' : "\033[1;33m" - colorcodes['bblue'] = monochrome_logs ? '' : "\033[1;34m" - colorcodes['bpurple'] = monochrome_logs ? '' : "\033[1;35m" - colorcodes['bcyan'] = monochrome_logs ? '' : "\033[1;36m" - colorcodes['bwhite'] = monochrome_logs ? '' : "\033[1;37m" - - // Underline - colorcodes['ublack'] = monochrome_logs ? '' : "\033[4;30m" - colorcodes['ured'] = monochrome_logs ? '' : "\033[4;31m" - colorcodes['ugreen'] = monochrome_logs ? '' : "\033[4;32m" - colorcodes['uyellow'] = monochrome_logs ? '' : "\033[4;33m" - colorcodes['ublue'] = monochrome_logs ? '' : "\033[4;34m" - colorcodes['upurple'] = monochrome_logs ? '' : "\033[4;35m" - colorcodes['ucyan'] = monochrome_logs ? '' : "\033[4;36m" - colorcodes['uwhite'] = monochrome_logs ? '' : "\033[4;37m" - - // High Intensity - colorcodes['iblack'] = monochrome_logs ? '' : "\033[0;90m" - colorcodes['ired'] = monochrome_logs ? '' : "\033[0;91m" - colorcodes['igreen'] = monochrome_logs ? '' : "\033[0;92m" - colorcodes['iyellow'] = monochrome_logs ? '' : "\033[0;93m" - colorcodes['iblue'] = monochrome_logs ? '' : "\033[0;94m" - colorcodes['ipurple'] = monochrome_logs ? '' : "\033[0;95m" - colorcodes['icyan'] = monochrome_logs ? '' : "\033[0;96m" - colorcodes['iwhite'] = monochrome_logs ? '' : "\033[0;97m" - - // Bold High Intensity - colorcodes['biblack'] = monochrome_logs ? '' : "\033[1;90m" - colorcodes['bired'] = monochrome_logs ? '' : "\033[1;91m" - colorcodes['bigreen'] = monochrome_logs ? '' : "\033[1;92m" - colorcodes['biyellow'] = monochrome_logs ? '' : "\033[1;93m" - colorcodes['biblue'] = monochrome_logs ? '' : "\033[1;94m" - colorcodes['bipurple'] = monochrome_logs ? '' : "\033[1;95m" - colorcodes['bicyan'] = monochrome_logs ? '' : "\033[1;96m" - colorcodes['biwhite'] = monochrome_logs ? '' : "\033[1;97m" - - return colorcodes - } - - // - // Does what is says on the tin - // - public static String dashedLine(monochrome_logs) { - Map colors = logColours(monochrome_logs) - return "-${colors.dim}----------------------------------------------------${colors.reset}-" - } - - // - // nf-core logo - // - public static String logo(workflow, monochrome_logs) { - Map colors = logColours(monochrome_logs) - String workflow_version = NfcoreTemplate.version(workflow) - String.format( - """\n - ${dashedLine(monochrome_logs)} - ${colors.purple} ${workflow.manifest.name} ${workflow_version}${colors.reset} - ${dashedLine(monochrome_logs)} - """.stripIndent() - ) - } -} diff --git a/lib/Utils.groovy b/lib/Utils.groovy deleted file mode 100755 index 8d030f4e..00000000 --- a/lib/Utils.groovy +++ /dev/null @@ -1,47 +0,0 @@ -// -// This file holds several Groovy functions that could be useful for any Nextflow pipeline -// - -import org.yaml.snakeyaml.Yaml - -class Utils { - - // - // When running with -profile conda, warn if channels have not been set-up appropriately - // - public static void checkCondaChannels(log) { - Yaml parser = new Yaml() - def channels = [] - try { - def config = parser.load("conda config --show channels".execute().text) - channels = config.channels - } catch(NullPointerException | IOException e) { - log.warn "Could not verify conda channel configuration." - return - } - - // Check that all channels are present - // This channel list is ordered by required channel priority. - def required_channels_in_order = ['conda-forge', 'bioconda', 'defaults'] - def channels_missing = ((required_channels_in_order as Set) - (channels as Set)) as Boolean - - // Check that they are in the right order - def channel_priority_violation = false - def n = required_channels_in_order.size() - for (int i = 0; i < n - 1; i++) { - channel_priority_violation |= !(channels.indexOf(required_channels_in_order[i]) < channels.indexOf(required_channels_in_order[i+1])) - } - - if (channels_missing | channel_priority_violation) { - log.warn "~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~\n" + - " There is a problem with your Conda configuration!\n\n" + - " You will need to set-up the conda-forge and bioconda channels correctly.\n" + - " Please refer to https://bioconda.github.io/\n" + - " The observed channel order is \n" + - " ${channels}\n" + - " but the following channel order is required:\n" + - " ${required_channels_in_order}\n" + - "~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~" - } - } -} diff --git a/lib/WorkflowMain.groovy b/lib/WorkflowMain.groovy deleted file mode 100755 index 3ae24dbf..00000000 --- a/lib/WorkflowMain.groovy +++ /dev/null @@ -1,63 +0,0 @@ -// -// This file holds several functions specific to the main.nf workflow in the qbic-pipelines/rnadeseq pipeline -// - -import nextflow.Nextflow - -class WorkflowMain { - - // - // Citation string for pipeline - // - public static String citation(workflow) { - return "If you use ${workflow.manifest.name} for your analysis please cite:\n\n" + - // TODO nf-core: Add Zenodo DOI for pipeline after first release - //"* The pipeline\n" + - //" https://doi.org/10.5281/zenodo.XXXXXXX\n\n" + - "* The nf-core framework\n" + - " https://doi.org/10.1038/s41587-020-0439-x\n\n" + - "* Software dependencies\n" + - " https://github.com/${workflow.manifest.name}/blob/master/CITATIONS.md" - } - - - // - // Validate parameters and print summary to screen - // - public static void initialise(workflow, params, log) { - - // Print workflow version and exit on --version - if (params.version) { - String workflow_version = NfcoreTemplate.version(workflow) - log.info "${workflow.manifest.name} ${workflow_version}" - System.exit(0) - } - - // Check that a -profile or Nextflow config has been provided to run the pipeline - NfcoreTemplate.checkConfigProvided(workflow, log) - - // Check that conda channels are set-up correctly - if (workflow.profile.tokenize(',').intersect(['conda', 'mamba']).size() >= 1) { - Utils.checkCondaChannels(log) - } - - // Check AWS batch settings - NfcoreTemplate.awsBatch(workflow, params) - - // Check input has been provided - if (!params.input) { - Nextflow.error("Please provide an input samplesheet to the pipeline e.g. '--input samplesheet.tsv'") - } - } - // - // Get attribute from genome config file e.g. fasta - // - public static Object getGenomeAttribute(params, attribute) { - if (params.genomes && params.genome && params.genomes.containsKey(params.genome)) { - if (params.genomes[ params.genome ].containsKey(attribute)) { - return params.genomes[ params.genome ][ attribute ] - } - } - return null - } -} diff --git a/lib/WorkflowRnadeseq.groovy b/lib/WorkflowRnadeseq.groovy deleted file mode 100755 index 7a1ae2bd..00000000 --- a/lib/WorkflowRnadeseq.groovy +++ /dev/null @@ -1,118 +0,0 @@ -// -// This file holds several functions specific to the workflow/rnadeseq.nf in the qbic-pipelines/rnadeseq pipeline -// - -import nextflow.Nextflow -import groovy.text.SimpleTemplateEngine - -class WorkflowRnadeseq { - - // - // Check and validate parameters - // - public static void initialise(params, log) { - - genomeExistsError(params, log) - - } - - // - // Get workflow summary for MultiQC - // - public static String paramsSummaryMultiqc(workflow, summary) { - String summary_section = '' - for (group in summary.keySet()) { - def group_params = summary.get(group) // This gets the parameters of that particular group - if (group_params) { - summary_section += "

    $group

    \n" - summary_section += "
    \n" - for (param in group_params.keySet()) { - summary_section += "
    $param
    ${group_params.get(param) ?: 'N/A'}
    \n" - } - summary_section += "
    \n" - } - } - - String yaml_file_text = "id: '${workflow.manifest.name.replace('/','-')}-summary'\n" - yaml_file_text += "description: ' - this information is collected when the pipeline is started.'\n" - yaml_file_text += "section_name: '${workflow.manifest.name} Workflow Summary'\n" - yaml_file_text += "section_href: 'https://github.com/${workflow.manifest.name}'\n" - yaml_file_text += "plot_type: 'html'\n" - yaml_file_text += "data: |\n" - yaml_file_text += "${summary_section}" - return yaml_file_text - } - - // - // Generate methods description for MultiQC - // - - public static String toolCitationText(params) { - - // TODO Optionally add in-text citation tools to this list. - // Can use ternary operators to dynamically construct based conditions, e.g. params["run_xyz"] ? "Tool (Foo et al. 2023)" : "", - // Uncomment function in methodsDescriptionText to render in MultiQC report - def citation_text = [ - "Tools used in the workflow included:", - "FastQC (Andrews 2010),", - "MultiQC (Ewels et al. 2016)", - "." - ].join(' ').trim() - - return citation_text - } - - public static String toolBibliographyText(params) { - - // TODO Optionally add bibliographic entries to this list. - // Can use ternary operators to dynamically construct based conditions, e.g. params["run_xyz"] ? "
  • Author (2023) Pub name, Journal, DOI
    • " : "", - // Uncomment function in methodsDescriptionText to render in MultiQC report - def reference_text = [ - "
  • Andrews S, (2010) FastQC, URL: https://www.bioinformatics.babraham.ac.uk/projects/fastqc/).
    • ", - "
  • Ewels, P., Magnusson, M., Lundin, S., & Käller, M. (2016). MultiQC: summarize analysis results for multiple tools and samples in a single report. Bioinformatics , 32(19), 3047–3048. doi: /10.1093/bioinformatics/btw354
    • " - ].join(' ').trim() - - return reference_text - } - - public static String methodsDescriptionText(run_workflow, mqc_methods_yaml, params) { - // Convert to a named map so can be used as with familar NXF ${workflow} variable syntax in the MultiQC YML file - def meta = [:] - meta.workflow = run_workflow.toMap() - meta["manifest_map"] = run_workflow.manifest.toMap() - - // Pipeline DOI - meta["doi_text"] = meta.manifest_map.doi ? "(doi: ${meta.manifest_map.doi})" : "" - meta["nodoi_text"] = meta.manifest_map.doi ? "": "
  • If available, make sure to update the text to include the Zenodo DOI of version of the pipeline used.
    • " - - // Tool references - meta["tool_citations"] = "" - meta["tool_bibliography"] = "" - - // TODO Only uncomment below if logic in toolCitationText/toolBibliographyText has been filled! - //meta["tool_citations"] = toolCitationText(params).replaceAll(", \\.", ".").replaceAll("\\. \\.", ".").replaceAll(", \\.", ".") - //meta["tool_bibliography"] = toolBibliographyText(params) - - - def methods_text = mqc_methods_yaml.text - - def engine = new SimpleTemplateEngine() - def description_html = engine.createTemplate(methods_text).make(meta) - - return description_html - } - - // - // Exit pipeline if incorrect --genome key provided - // - private static void genomeExistsError(params, log) { - if (params.genomes && params.genome && !params.genomes.containsKey(params.genome)) { - def error_string = "~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~\n" + - " Genome '${params.genome}' not found in any config files provided to the pipeline.\n" + - " Currently, the available genome keys are:\n" + - " ${params.genomes.keySet().join(", ")}\n" + - "~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~" - Nextflow.error(error_string) - } - } -} diff --git a/lib/nfcore_external_java_deps.jar b/lib/nfcore_external_java_deps.jar deleted file mode 100644 index 805c8bb5e4fd43a12a5891eea5a68788309629b0..0000000000000000000000000000000000000000 GIT binary patch literal 0 HcmV?d00001 literal 2291171 zcma%i1CS<7mu}m(-Ea4_?dhK8v~AnAZQHhO+qP}n-E-&Lf8*Yb{bM(7MpZ;s){{|L znGpxiIY(9k6buar3JMCSNkdW|=)W{*ATS^)VMTsgF=-L{?{Od?P#{?eNT`1hApZ%I z{U0Zz{YCuW$x{5%Vj{u{igZ#U4^op8k`lCZbMO+hRMV3)4e|_&%)3YSbmCGpVp4NX zni_mKLGakt&P12{r^KCD0GPa9sIuy`fu+agt?8O-hY9i{WloGe}y@kSsDEY_Fud9 z-?2ab4J)MQXe4E3Xl-U<>PRMQZ)0m@@90KnV5#Te5Tz`pvZ0Lj4F<9%Flh(`Qb;Tn zMV!iFmH{S^fF*j#MCKRUd`7m>9||*;WE32I_6pg4-Q1C#ke$$aVmP-TtfJ+j+N9;Q z>bxN=bCVQkg$Jzty71(+~A9}ZfANuYIVnTdW!??Sj)Ti-Qpk8XgPDab8pk2QD&dwUl=iFhd1c$C2uMVagr zWc@z9s$#g__{3?VGb#)^fSNp^lZh 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WorkflowMain.getGenomeAttribute(params, 'gtf') + params.gtf = getGenomeAttribute('gtf') if (!params.gtf) { exit 1, 'It seems that for your genome, no gtf file is defined. Please provide a gtf with the --gtf parameter or open a github issue: https://github.com/qbic-pipelines/rnadeseq/issues' } @@ -40,44 +40,27 @@ if (params.input_type in ["rsem", "salmon"]) { if (params.run_pathway_analysis) { if (!params.genome && !params.organism) { exit 1, 'Please provide either genome or organism!' } else if (!params.organism) { - params.organism = WorkflowMain.getGenomeAttribute(params, 'organism') + params.organism = getGenomeAttribute('organism') if (!params.organism) { exit 1, 'It seems that for your genome, no organism is defined. Please provide the organism with the --organism parameter or open a github issue: https://github.com/qbic-pipelines/rnadeseq/issues' } } if (!params.genome && !params.species_library) { exit 1, 'Please provide either genome or species_library!' } else if (!params.species_library) { - params.species_library = WorkflowMain.getGenomeAttribute(params, 'species_library') + params.species_library = getGenomeAttribute('species_library') if (!params.species_library) { exit 1, 'It seems that for your genome, no species_library is defined. Please provide the library with the --species_library parameter or open a github issue: https://github.com/qbic-pipelines/rnadeseq/issues' } } if (!params.genome && !params.keytype) { exit 1, 'Please provide either genome or keytype!' } else if (!params.keytype) { - params.keytype = WorkflowMain.getGenomeAttribute(params, 'keytype') + params.keytype = getGenomeAttribute('keytype') if (!params.keytype) { exit 1, 'It seems that for your genome, no keytype is defined. Please provide the keytype with the --keytype parameter or open a github issue: https://github.com/qbic-pipelines/rnadeseq/issues' } } } -/* -~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ - PRINT PARAMETER SUMMARY -~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ -*/ - -// Print help message if needed -if (params.help) { - def logo = NfcoreTemplate.logo(workflow, params.monochrome_logs) - def citation = '\n' + WorkflowMain.citation(workflow) + '\n' - def String command = "nextflow run ${workflow.manifest.name} --input 'samplesheet.tsv' --gene_counts 'merged_gene_counts.txt' --input 'sample_preparations.tsv' --model 'linear_model.txt' -profile docker" - log.info logo + paramsHelp(command) + citation + NfcoreTemplate.dashedLine(params.monochrome_logs) - System.exit(0) -} - -WorkflowMain.initialise(workflow, params, log) - /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ NAMED WORKFLOW FOR PIPELINE diff --git a/nextflow.config b/nextflow.config index 9696be8a..3b8f5a4b 100644 --- a/nextflow.config +++ b/nextflow.config @@ -86,6 +86,9 @@ params { max_cpus = 16 max_time = '240.h' + // Schema validation default options (only those necessary for linting; rnadeseq does not use the validation plugin) + validationSchemaIgnoreParams = 'genomes,igenomes_base' + validationShowHiddenParams = false } // Load base.config by default for all pipelines diff --git a/nextflow_schema.json b/nextflow_schema.json index b5b65217..b1b923d6 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -125,6 +125,7 @@ }, "custom_gmt": { "type": "string", + "default": "NO_FILE3", "description": "Path to custom GMT file for gost query, this allows e.g. to query older versions of databases for pathway analysis, not necessary if --skip_pathway_analysis = true. If --datasources is set, the GMT file will be filtered for these datasources." }, "set_background": { @@ -134,6 +135,7 @@ }, "custom_background": { "type": "string", + "default": "NO_FILE7", "description": "Path to custom background TXT file for gost query containing a gene ID in each line, not necessary if --skip_pathway_analysis = true or --set_background = false." }, "min_DEG_pathway": { @@ -169,15 +171,17 @@ }, "multiqc": { "type": "string", + "default": "NO_FILE4", "description": "multiqc.zip folder containing the multiQC plots and report." }, "quote": { "type": "string", + "default": "NO_FILE5", "description": "Quote file to link in the report." }, "software_versions": { "type": "string", - "default": "None", + "default": "NO_FILE6", "description": "CSV/YML containing nf-core/rnaseq software versions." } } @@ -190,7 +194,6 @@ "properties": { "genome": { "type": "string", - "default": "None", "description": "Which genome to use for analysis, e.g. GRCh37; see /conf/igenomes.config for which genomes are available." }, "igenomes_base": { @@ -225,7 +228,6 @@ "properties": { "publish_dir_mode": { "type": "string", - "default": "copy", "description": "Method used to save pipeline results to output directory.", "help_text": "The Nextflow `publishDir` option specifies which intermediate files should be saved to the output directory. This option tells the pipeline what method should be used to move these files. See [Nextflow docs](https://www.nextflow.io/docs/latest/process.html#publishdir) for details.", "fa_icon": "fas fa-copy", @@ -248,8 +250,7 @@ "properties": { "email": { "type": "string", - "description": "Set this parameter to your e-mail address to get a summary e-mail with details of the run sent to you when the workflow exits", - "default": "false" + "description": "Set this parameter to your e-mail address to get a summary e-mail with details of the run sent to you when the workflow exits" }, "email_on_fail": { "type": "string", @@ -295,19 +296,16 @@ }, "config_profile_description": { "type": "string", - "default": "false", "description": "Institutional config description.", "hidden": true }, "config_profile_contact": { "type": "string", - "default": "false", "description": "Institutional config contact information.", "hidden": true }, "config_profile_url": { "type": "string", - "default": "false", "description": "Institutional config URL link.", "hidden": true }, diff --git a/workflows/rnadeseq.nf b/workflows/rnadeseq.nf index c2941c0f..4ba022c8 100644 --- a/workflows/rnadeseq.nf +++ b/workflows/rnadeseq.nf @@ -4,14 +4,11 @@ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ -// Validate input parameters -WorkflowRnadeseq.initialise(params, log) - // TODO nf-core: Add all file path parameters for the pipeline to the list below // Check input path parameters to see if they exist def checkPathParamList = [ - params.input, params.model, - ] + params.input, params.model +] for (param in checkPathParamList) { if (param) { file(param, checkIfExists: true) } } // Check mandatory parameters @@ -36,31 +33,12 @@ ch_custom_background = Channel.fromPath(params.custom_background) ch_proj_summary_file = Channel.fromPath(params.project_summary) ch_softwareversions_file = Channel.fromPath(params.software_versions) -/* -~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ - PRINT PARAMS SUMMARY -~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ -*/ - -include { paramsSummaryLog; paramsSummaryMap } from 'plugin/nf-validation' - -def logo = NfcoreTemplate.logo(workflow, params.monochrome_logs) -def citation = '\n' + WorkflowMain.citation(workflow) + '\n' -def summary_params = paramsSummaryMap(workflow) - -// Print parameter summary log to screen -log.info logo + paramsSummaryLog(workflow) + citation - -WorkflowRnadeseq.initialise(params, log) - /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ IMPORT LOCAL MODULES/SUBWORKFLOWS ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ -// -// MODULE: Loaded from modules/local/ -// + include { REPORT } from '../modules/local/report' /* @@ -69,9 +47,6 @@ include { REPORT } from '../modules/local/report' ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ -// -// MODULE: Installed directly from nf-core/modules -// include { CUSTOM_DUMPSOFTWAREVERSIONS } from '../modules/nf-core/custom/dumpsoftwareversions/main' /* From 1630114541e18d9204222ad9ede5e6507fc99ef5 Mon Sep 17 00:00:00 2001 From: WackerO Date: Thu, 16 May 2024 09:42:29 +0200 Subject: [PATCH 54/73] fix linting --- .nf-core.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/.nf-core.yml b/.nf-core.yml index f763727e..264c6e65 100644 --- a/.nf-core.yml +++ b/.nf-core.yml @@ -20,4 +20,4 @@ lint: - params.custom_config_base multiqc_config: - - report_comment # otherwise throws error: multiqc_config: 'assets/multiqc_config.yml' does not contain a matching 'report_comment'. + - report_comment # otherwise throws error: multiqc_config: 'assets/multiqc_config.yml' does not contain a matching 'report_comment'. From b704437d3c4146005efd0fffd73f219303927640 Mon Sep 17 00:00:00 2001 From: WackerO Date: Thu, 16 May 2024 09:50:44 +0200 Subject: [PATCH 55/73] prettier --- .nf-core.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/.nf-core.yml b/.nf-core.yml index 264c6e65..01625cf3 100644 --- a/.nf-core.yml +++ b/.nf-core.yml @@ -17,7 +17,7 @@ lint: - manifest.name - manifest.homePage - config_defaults: - - params.custom_config_base + - params.custom_config_base multiqc_config: - report_comment # otherwise throws error: multiqc_config: 'assets/multiqc_config.yml' does not contain a matching 'report_comment'. From fa909d755cab58f09de8d0ece3b9723d7c6e5964 Mon Sep 17 00:00:00 2001 From: WackerO Date: Thu, 16 May 2024 10:56:21 +0200 Subject: [PATCH 56/73] Removed TODOs, removed nf-validation plugin import, changed 1x nf-core-rnadeseq to qbic-pipelines-rnadeseq --- .github/workflows/awsfulltest.yml | 3 --- assets/methods_description_template.yml | 1 - assets/multiqc_config.yml | 4 ++-- conf/base.config | 2 -- nextflow.config | 10 ---------- .../local/utils_nfcore_rnadeseq_pipeline/main.nf | 7 ------- workflows/rnadeseq.nf | 1 - 7 files changed, 2 insertions(+), 26 deletions(-) diff --git a/.github/workflows/awsfulltest.yml b/.github/workflows/awsfulltest.yml index 63788533..029731bf 100644 --- a/.github/workflows/awsfulltest.yml +++ b/.github/workflows/awsfulltest.yml @@ -15,9 +15,6 @@ jobs: steps: - name: Launch workflow via Seqera Platform uses: seqeralabs/action-tower-launch@v2 - # TODO nf-core: You can customise AWS full pipeline tests as required - # Add full size test data (but still relatively small datasets for few samples) - # on the `test_full.config` test runs with only one set of parameters with: workspace_id: ${{ secrets.TOWER_WORKSPACE_ID }} access_token: ${{ secrets.TOWER_ACCESS_TOKEN }} diff --git a/assets/methods_description_template.yml b/assets/methods_description_template.yml index 6212134d..f5f5bfe4 100644 --- a/assets/methods_description_template.yml +++ b/assets/methods_description_template.yml @@ -3,7 +3,6 @@ description: "Suggested text and references to use when describing pipeline usag section_name: "qbic-pipelines/rnadeseq Methods Description" section_href: "https://github.com/qbic-pipelines/rnadeseq" plot_type: "html" -## TODO nf-core: Update the HTML below to your preferred methods description, e.g. add publication citation for this pipeline ## You inject any metadata in the Nextflow '${workflow}' object data: |

    Methods

    diff --git a/assets/multiqc_config.yml b/assets/multiqc_config.yml index 439b820e..d3a5bdca 100644 --- a/assets/multiqc_config.yml +++ b/assets/multiqc_config.yml @@ -1,9 +1,9 @@ report_comment: > This report has been generated by the qbic-pipelines/rnadeseq analysis pipeline. For information about how to interpret these results, please see the - documentation. + documentation. report_section_order: - "nf-core-rnadeseq-methods-description": + "qbic-pipelines-rnadeseq-methods-description": order: -1000 software_versions: order: -1001 diff --git a/conf/base.config b/conf/base.config index 6ce7a323..e68d4caf 100644 --- a/conf/base.config +++ b/conf/base.config @@ -10,7 +10,6 @@ process { - // TODO nf-core: Check the defaults for all processes cpus = { check_max( 1 * task.attempt, 'cpus' ) } memory = { check_max( 6.GB * task.attempt, 'memory' ) } time = { check_max( 4.h * task.attempt, 'time' ) } @@ -24,7 +23,6 @@ process { // These labels are used and recognised by default in DSL2 files hosted on nf-core/modules. // If possible, it would be nice to keep the same label naming convention when // adding in your local modules too. - // TODO nf-core: Customise requirements for specific processes. // See https://www.nextflow.io/docs/latest/config.html#config-process-selectors withLabel:process_single { cpus = { check_max( 1 , 'cpus' ) } diff --git a/nextflow.config b/nextflow.config index 3b8f5a4b..f644b6fe 100644 --- a/nextflow.config +++ b/nextflow.config @@ -6,11 +6,6 @@ ---------------------------------------------------------------------------------------- */ -// Pin nf-validation to prevent errors from new versions -plugins { - id 'nf-validation@1.1.3' -} - // Global default params, used in configs // params marked with //* exist for the sake of completeness only; // enabling them prevents igenomes from working @@ -228,11 +223,6 @@ docker.registry = 'quay.io' podman.registry = 'quay.io' singularity.registry = 'quay.io' -// Nextflow plugins -plugins { - id 'nf-validation@1.1.3' // Validation of pipeline parameters and creation of an input channel from a sample sheet -} - // Load igenomes.config if required if (!params.igenomes_ignore) { includeConfig 'conf/igenomes.config' diff --git a/subworkflows/local/utils_nfcore_rnadeseq_pipeline/main.nf b/subworkflows/local/utils_nfcore_rnadeseq_pipeline/main.nf index 95c433cc..ca9163bd 100644 --- a/subworkflows/local/utils_nfcore_rnadeseq_pipeline/main.nf +++ b/subworkflows/local/utils_nfcore_rnadeseq_pipeline/main.nf @@ -202,7 +202,6 @@ def genomeExistsError() { // Generate methods description for MultiQC // def toolCitationText() { - // TODO nf-core: Optionally add in-text citation tools to this list. // Can use ternary operators to dynamically construct based conditions, e.g. params["run_xyz"] ? "Tool (Foo et al. 2023)" : "", // Uncomment function in methodsDescriptionText to render in MultiQC report def citation_text = [ @@ -216,7 +215,6 @@ def toolCitationText() { } def toolBibliographyText() { - // TODO nf-core: Optionally add bibliographic entries to this list. // Can use ternary operators to dynamically construct based conditions, e.g. params["run_xyz"] ? "
  • Author (2023) Pub name, Journal, DOI
    • " : "", // Uncomment function in methodsDescriptionText to render in MultiQC report def reference_text = [ @@ -249,11 +247,6 @@ def methodsDescriptionText(mqc_methods_yaml) { meta["tool_citations"] = "" meta["tool_bibliography"] = "" - // TODO nf-core: Only uncomment below if logic in toolCitationText/toolBibliographyText has been filled! - // meta["tool_citations"] = toolCitationText().replaceAll(", \\.", ".").replaceAll("\\. \\.", ".").replaceAll(", \\.", ".") - // meta["tool_bibliography"] = toolBibliographyText() - - def methods_text = mqc_methods_yaml.text def engine = new groovy.text.SimpleTemplateEngine() diff --git a/workflows/rnadeseq.nf b/workflows/rnadeseq.nf index 4ba022c8..a47b3a0c 100644 --- a/workflows/rnadeseq.nf +++ b/workflows/rnadeseq.nf @@ -4,7 +4,6 @@ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ -// TODO nf-core: Add all file path parameters for the pipeline to the list below // Check input path parameters to see if they exist def checkPathParamList = [ params.input, params.model From 5fa7b2981b18899fb28d6c15f5df5300eb035f93 Mon Sep 17 00:00:00 2001 From: WackerO Date: Thu, 16 May 2024 11:12:15 +0200 Subject: [PATCH 57/73] added lib entry to nf-core.yml to skip error --- .nf-core.yml | 3 +++ 1 file changed, 3 insertions(+) diff --git a/.nf-core.yml b/.nf-core.yml index 01625cf3..4ce60765 100644 --- a/.nf-core.yml +++ b/.nf-core.yml @@ -13,6 +13,9 @@ lint: - docs/README.md - LICENSE + files_exist: + - lib/nfcore_external_java_deps.jar + nextflow_config: - manifest.name - manifest.homePage From 91d9b3d2b6c2d711ffe74c210d9e23edff9b9df1 Mon Sep 17 00:00:00 2001 From: WackerO Date: Thu, 16 May 2024 12:52:10 +0200 Subject: [PATCH 58/73] removed some unnecessary code --- .github/workflows/release-announcements.yml | 75 ------ assets/samplesheet.csv | 3 - assets/schema_input.json | 33 --- subworkflows/local/input_check.nf | 44 ---- .../utils_nfcore_rnadeseq_pipeline/main.nf | 240 ------------------ 5 files changed, 395 deletions(-) delete mode 100644 .github/workflows/release-announcements.yml delete mode 100644 assets/samplesheet.csv delete mode 100644 assets/schema_input.json delete mode 100644 subworkflows/local/input_check.nf diff --git a/.github/workflows/release-announcements.yml b/.github/workflows/release-announcements.yml deleted file mode 100644 index 03ecfcf7..00000000 --- a/.github/workflows/release-announcements.yml +++ /dev/null @@ -1,75 +0,0 @@ -name: release-announcements -# Automatic release toot and tweet anouncements -on: - release: - types: [published] - workflow_dispatch: - -jobs: - toot: - runs-on: ubuntu-latest - steps: - - name: get topics and convert to hashtags - id: get_topics - run: | - echo "topics=$(curl -s https://nf-co.re/pipelines.json | jq -r '.remote_workflows[] | select(.full_name == "${{ github.repository }}") | .topics[]' | awk '{print "#"$0}' | tr '\n' ' ')" >> $GITHUB_OUTPUT - - - uses: rzr/fediverse-action@master - with: - access-token: ${{ secrets.MASTODON_ACCESS_TOKEN }} - host: "mstdn.science" # custom host if not "mastodon.social" (default) - # GitHub event payload - # https://docs.github.com/en/developers/webhooks-and-events/webhooks/webhook-events-and-payloads#release - message: | - Pipeline release! ${{ github.repository }} v${{ github.event.release.tag_name }} - ${{ github.event.release.name }}! - - Please see the changelog: ${{ github.event.release.html_url }} - - ${{ steps.get_topics.outputs.topics }} #nfcore #openscience #nextflow #bioinformatics - - send-tweet: - runs-on: ubuntu-latest - - steps: - - uses: actions/setup-python@82c7e631bb3cdc910f68e0081d67478d79c6982d # v5 - with: - python-version: "3.10" - - name: Install dependencies - run: pip install tweepy==4.14.0 - - name: Send tweet - shell: python - run: | - import os - import tweepy - - client = tweepy.Client( - access_token=os.getenv("TWITTER_ACCESS_TOKEN"), - access_token_secret=os.getenv("TWITTER_ACCESS_TOKEN_SECRET"), - consumer_key=os.getenv("TWITTER_CONSUMER_KEY"), - consumer_secret=os.getenv("TWITTER_CONSUMER_SECRET"), - ) - tweet = os.getenv("TWEET") - client.create_tweet(text=tweet) - env: - TWEET: | - Pipeline release! ${{ github.repository }} v${{ github.event.release.tag_name }} - ${{ github.event.release.name }}! - - Please see the changelog: ${{ github.event.release.html_url }} - TWITTER_CONSUMER_KEY: ${{ secrets.TWITTER_CONSUMER_KEY }} - TWITTER_CONSUMER_SECRET: ${{ secrets.TWITTER_CONSUMER_SECRET }} - TWITTER_ACCESS_TOKEN: ${{ secrets.TWITTER_ACCESS_TOKEN }} - TWITTER_ACCESS_TOKEN_SECRET: ${{ secrets.TWITTER_ACCESS_TOKEN_SECRET }} - - bsky-post: - runs-on: ubuntu-latest - steps: - - uses: zentered/bluesky-post-action@80dbe0a7697de18c15ad22f4619919ceb5ccf597 # v0.1.0 - with: - post: | - Pipeline release! ${{ github.repository }} v${{ github.event.release.tag_name }} - ${{ github.event.release.name }}! - - Please see the changelog: ${{ github.event.release.html_url }} - env: - BSKY_IDENTIFIER: ${{ secrets.BSKY_IDENTIFIER }} - BSKY_PASSWORD: ${{ secrets.BSKY_PASSWORD }} - # diff --git a/assets/samplesheet.csv b/assets/samplesheet.csv deleted file mode 100644 index 5f653ab7..00000000 --- a/assets/samplesheet.csv +++ /dev/null @@ -1,3 +0,0 @@ -sample,fastq_1,fastq_2 -SAMPLE_PAIRED_END,/path/to/fastq/files/AEG588A1_S1_L002_R1_001.fastq.gz,/path/to/fastq/files/AEG588A1_S1_L002_R2_001.fastq.gz -SAMPLE_SINGLE_END,/path/to/fastq/files/AEG588A4_S4_L003_R1_001.fastq.gz, diff --git a/assets/schema_input.json b/assets/schema_input.json deleted file mode 100644 index 824a9ce8..00000000 --- a/assets/schema_input.json +++ /dev/null @@ -1,33 +0,0 @@ -{ - "$schema": "http://json-schema.org/draft-07/schema", - "$id": "https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/master/assets/schema_input.json", - "title": "qbic-pipelines/rnadeseq pipeline - params.input schema", - "description": "Schema for the file provided with params.input", - "type": "array", - "items": { - "type": "object", - "properties": { - "sample": { - "type": "string", - "pattern": "^\\S+$", - "errorMessage": "Sample name must be provided and cannot contain spaces", - "meta": ["id"] - }, - "fastq_1": { - "type": "string", - "format": "file-path", - "exists": true, - "pattern": "^\\S+\\.f(ast)?q\\.gz$", - "errorMessage": "FastQ file for reads 1 must be provided, cannot contain spaces and must have extension '.fq.gz' or '.fastq.gz'" - }, - "fastq_2": { - "type": "string", - "format": "file-path", - "exists": true, - "pattern": "^\\S+\\.f(ast)?q\\.gz$", - "errorMessage": "FastQ file for reads 2 cannot contain spaces and must have extension '.fq.gz' or '.fastq.gz'" - } - }, - "required": ["sample", "fastq_1"] - } -} diff --git a/subworkflows/local/input_check.nf b/subworkflows/local/input_check.nf deleted file mode 100644 index 2fdd1e4f..00000000 --- a/subworkflows/local/input_check.nf +++ /dev/null @@ -1,44 +0,0 @@ -// -// Check input samplesheet and get read channels -// - -include { SAMPLESHEET_CHECK } from '../../modules/local/samplesheet_check' - -workflow INPUT_CHECK { - take: - samplesheet // file: /path/to/samplesheet.csv - - main: - SAMPLESHEET_CHECK ( samplesheet ) - .csv - .splitCsv ( header:true, sep:',' ) - .map { create_fastq_channel(it) } - .set { reads } - - emit: - reads // channel: [ val(meta), [ reads ] ] - software_versions = SAMPLESHEET_CHECK.out.versions // channel: [ software_versions.yml ] -} - -// Function to get list of [ meta, [ fastq_1, fastq_2 ] ] -def create_fastq_channel(LinkedHashMap row) { - // create meta map - def meta = [:] - meta.id = row.sample - meta.single_end = row.single_end.toBoolean() - - // add path(s) of the fastq file(s) to the meta map - def fastq_meta = [] - if (!file(row.fastq_1).exists()) { - exit 1, "ERROR: Please check input samplesheet -> Read 1 FastQ file does not exist!\n${row.fastq_1}" - } - if (meta.single_end) { - fastq_meta = [ meta, [ file(row.fastq_1) ] ] - } else { - if (!file(row.fastq_2).exists()) { - exit 1, "ERROR: Please check input samplesheet -> Read 2 FastQ file does not exist!\n${row.fastq_2}" - } - fastq_meta = [ meta, [ file(row.fastq_1), file(row.fastq_2) ] ] - } - return fastq_meta -} diff --git a/subworkflows/local/utils_nfcore_rnadeseq_pipeline/main.nf b/subworkflows/local/utils_nfcore_rnadeseq_pipeline/main.nf index ca9163bd..eb37c27b 100644 --- a/subworkflows/local/utils_nfcore_rnadeseq_pipeline/main.nf +++ b/subworkflows/local/utils_nfcore_rnadeseq_pipeline/main.nf @@ -1,178 +1,9 @@ -// -// Subworkflow with functionality specific to the qbic-pipelines/rnadeseq pipeline -// - -/* -~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ - IMPORT FUNCTIONS / MODULES / SUBWORKFLOWS -~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ -*/ - -include { UTILS_NFVALIDATION_PLUGIN } from '../../nf-core/utils_nfvalidation_plugin' -include { paramsSummaryMap } from 'plugin/nf-validation' -include { fromSamplesheet } from 'plugin/nf-validation' -include { UTILS_NEXTFLOW_PIPELINE } from '../../nf-core/utils_nextflow_pipeline' -include { completionEmail } from '../../nf-core/utils_nfcore_pipeline' -include { completionSummary } from '../../nf-core/utils_nfcore_pipeline' -include { dashedLine } from '../../nf-core/utils_nfcore_pipeline' -include { nfCoreLogo } from '../../nf-core/utils_nfcore_pipeline' -include { imNotification } from '../../nf-core/utils_nfcore_pipeline' -include { UTILS_NFCORE_PIPELINE } from '../../nf-core/utils_nfcore_pipeline' -include { workflowCitation } from '../../nf-core/utils_nfcore_pipeline' - -/* -======================================================================================== - SUBWORKFLOW TO INITIALISE PIPELINE -======================================================================================== -*/ - -workflow PIPELINE_INITIALISATION { - - take: - version // boolean: Display version and exit - help // boolean: Display help text - validate_params // boolean: Boolean whether to validate parameters against the schema at runtime - monochrome_logs // boolean: Do not use coloured log outputs - nextflow_cli_args // array: List of positional nextflow CLI args - outdir // string: The output directory where the results will be saved - input // string: Path to input samplesheet - - main: - - ch_versions = Channel.empty() - - // - // Print version and exit if required and dump pipeline parameters to JSON file - // - UTILS_NEXTFLOW_PIPELINE ( - version, - true, - outdir, - workflow.profile.tokenize(',').intersect(['conda', 'mamba']).size() >= 1 - ) - - // - // Validate parameters and generate parameter summary to stdout - // - pre_help_text = nfCoreLogo(monochrome_logs) - post_help_text = '\n' + workflowCitation() + '\n' + dashedLine(monochrome_logs) - def String workflow_command = "nextflow run ${workflow.manifest.name} -profile --input samplesheet.csv --outdir " - UTILS_NFVALIDATION_PLUGIN ( - help, - workflow_command, - pre_help_text, - post_help_text, - validate_params, - "nextflow_schema.json" - ) - - // - // Check config provided to the pipeline - // - UTILS_NFCORE_PIPELINE ( - nextflow_cli_args - ) - // - // Custom validation for pipeline parameters - // - validateInputParameters() - - // - // Create channel from input file provided through params.input - // - Channel - .fromSamplesheet("input") - .map { - meta, fastq_1, fastq_2 -> - if (!fastq_2) { - return [ meta.id, meta + [ single_end:true ], [ fastq_1 ] ] - } else { - return [ meta.id, meta + [ single_end:false ], [ fastq_1, fastq_2 ] ] - } - } - .groupTuple() - .map { - validateInputSamplesheet(it) - } - .map { - meta, fastqs -> - return [ meta, fastqs.flatten() ] - } - .set { ch_samplesheet } - - emit: - samplesheet = ch_samplesheet - versions = ch_versions -} - -/* -======================================================================================== - SUBWORKFLOW FOR PIPELINE COMPLETION -======================================================================================== -*/ - -workflow PIPELINE_COMPLETION { - - take: - email // string: email address - email_on_fail // string: email address sent on pipeline failure - plaintext_email // boolean: Send plain-text email instead of HTML - outdir // path: Path to output directory where results will be published - monochrome_logs // boolean: Disable ANSI colour codes in log output - hook_url // string: hook URL for notifications - multiqc_report // string: Path to MultiQC report - - main: - - summary_params = paramsSummaryMap(workflow, parameters_schema: "nextflow_schema.json") - - // - // Completion email and summary - // - workflow.onComplete { - if (email || email_on_fail) { - completionEmail(summary_params, email, email_on_fail, plaintext_email, outdir, monochrome_logs, multiqc_report.toList()) - } - - completionSummary(monochrome_logs) - - if (hook_url) { - imNotification(summary_params, hook_url) - } - } - - workflow.onError { - log.error "Pipeline failed. Please refer to troubleshooting docs: https://nf-co.re/docs/usage/troubleshooting" - } -} - /* ======================================================================================== FUNCTIONS ======================================================================================== */ // -// Check and validate pipeline parameters -// -def validateInputParameters() { - genomeExistsError() -} - -// -// Validate channels from input samplesheet -// -def validateInputSamplesheet(input) { - def (metas, fastqs) = input[1..2] - - // Check that multiple runs of the same sample are of the same datatype i.e. single-end / paired-end - def endedness_ok = metas.collect{ it.single_end }.unique().size == 1 - if (!endedness_ok) { - error("Please check input samplesheet -> Multiple runs of a sample must be of the same datatype i.e. single-end or paired-end: ${metas[0].id}") - } - - return [ metas[0], fastqs ] -} -// // Get attribute from genome config file e.g. fasta // def getGenomeAttribute(attribute) { @@ -183,74 +14,3 @@ def getGenomeAttribute(attribute) { } return null } - -// -// Exit pipeline if incorrect --genome key provided -// -def genomeExistsError() { - if (params.genomes && params.genome && !params.genomes.containsKey(params.genome)) { - def error_string = "~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~\n" + - " Genome '${params.genome}' not found in any config files provided to the pipeline.\n" + - " Currently, the available genome keys are:\n" + - " ${params.genomes.keySet().join(", ")}\n" + - "~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~" - error(error_string) - } -} - -// -// Generate methods description for MultiQC -// -def toolCitationText() { - // Can use ternary operators to dynamically construct based conditions, e.g. params["run_xyz"] ? "Tool (Foo et al. 2023)" : "", - // Uncomment function in methodsDescriptionText to render in MultiQC report - def citation_text = [ - "Tools used in the workflow included:", - "FastQC (Andrews 2010),", - "MultiQC (Ewels et al. 2016)", - "." - ].join(' ').trim() - - return citation_text -} - -def toolBibliographyText() { - // Can use ternary operators to dynamically construct based conditions, e.g. params["run_xyz"] ? "
  • Author (2023) Pub name, Journal, DOI
    • " : "", - // Uncomment function in methodsDescriptionText to render in MultiQC report - def reference_text = [ - "
  • Andrews S, (2010) FastQC, URL: https://www.bioinformatics.babraham.ac.uk/projects/fastqc/).
    • ", - "
  • Ewels, P., Magnusson, M., Lundin, S., & Käller, M. (2016). MultiQC: summarize analysis results for multiple tools and samples in a single report. Bioinformatics , 32(19), 3047–3048. doi: /10.1093/bioinformatics/btw354
    • " - ].join(' ').trim() - - return reference_text -} - -def methodsDescriptionText(mqc_methods_yaml) { - // Convert to a named map so can be used as with familar NXF ${workflow} variable syntax in the MultiQC YML file - def meta = [:] - meta.workflow = workflow.toMap() - meta["manifest_map"] = workflow.manifest.toMap() - - // Pipeline DOI - if (meta.manifest_map.doi) { - // Using a loop to handle multiple DOIs - // Removing `https://doi.org/` to handle pipelines using DOIs vs DOI resolvers - // Removing ` ` since the manifest.doi is a string and not a proper list - def temp_doi_ref = "" - String[] manifest_doi = meta.manifest_map.doi.tokenize(",") - for (String doi_ref: manifest_doi) temp_doi_ref += "(doi: ${doi_ref.replace("https://doi.org/", "").replace(" ", "")}), " - meta["doi_text"] = temp_doi_ref.substring(0, temp_doi_ref.length() - 2) - } else meta["doi_text"] = "" - meta["nodoi_text"] = meta.manifest_map.doi ? "" : "
  • If available, make sure to update the text to include the Zenodo DOI of version of the pipeline used.
    • " - - // Tool references - meta["tool_citations"] = "" - meta["tool_bibliography"] = "" - - def methods_text = mqc_methods_yaml.text - - def engine = new groovy.text.SimpleTemplateEngine() - def description_html = engine.createTemplate(methods_text).make(meta) - - return description_html.toString() -} From 8b93f6240f480b66e4f4cf5daa8fddaf78f5bdfe Mon Sep 17 00:00:00 2001 From: WackerO Date: Thu, 23 May 2024 09:08:05 +0200 Subject: [PATCH 59/73] add logic to better check which multiqc version is provided --- assets/RNAseq_report.Rmd | 59 ++++++++++++++++++++++++++-------------- 1 file changed, 38 insertions(+), 21 deletions(-) diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index 8d79f7f8..2042c5e2 100644 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -371,7 +371,7 @@ metadata$Secondary.Name <- gsub(" ; ", "_", metadata$Secondary.Name) metadata$Secondary.Name <- gsub(" ", "_", metadata$Secondary.Name) metadata$sampleName = paste(row.names(metadata),metadata$Secondary.Name,sep="_") row.names(metadata) = metadata$sampleName -metadata_save <- metadata +metadata_save <- metadata # save copy so that original metadata object can be changed if (params$input_type == "smrnaseq") { @@ -872,12 +872,46 @@ for (i in resultsNames(cds)[-1]) { # Check first if a new or old multiqc file was provided if (file.exists(paste0(wd,"/QC/multiqc_data/multiqc_general_stats.txt"))) { mqc_stats <- read.table(file = paste0(wd,"/QC/multiqc_data/multiqc_general_stats.txt"), header=TRUE, sep="\t") - mqc_version <- 'old_mqc' + columns <- c( + "Sample", + "FastQC_mqc.generalstats.fastqc.total_sequences", + "FastQC_mqc.generalstats.fastqc.percent_duplicates", + "FastQC_mqc.generalstats.fastqc.percent_gc", + "Cutadapt_mqc.generalstats.cutadapt.percent_trimmed", + "STAR_mqc.generalstats.star.uniquely_mapped_percent", + "featureCounts_mqc.generalstats.featurecounts.percent_assigned" + ) + if (all(columns %in% colnames(mqc_stats))) { + mqc_version <- 'old_mqc' + } else if (file.exists(paste0(wd,"/QC/multiqc_data/multiqc_samtools_stats.txt"))) { + mqc_stats <- read.table(file = paste0(wd,"/QC/multiqc_data/multiqc_samtools_stats.txt"), header=TRUE, sep="\t") + columns <- c( + "Sample", + "raw_total_sequences", + "reads_mapped_percent", + "reads_duplicated_percent" + ) + if (all(columns %in% colnames(mqc_stats))) { + mqc_version <- 'new_mqc' + } else { + stop("Could not find a suitable multiqc table; please provide a correct multiqc.zip file or omit the parameter --multiqc altogether.") + } + } } else if (file.exists(paste0(wd,"/QC/multiqc_data/multiqc_samtools_stats.txt"))) { mqc_stats <- read.table(file = paste0(wd,"/QC/multiqc_data/multiqc_samtools_stats.txt"), header=TRUE, sep="\t") - mqc_version <- 'new_mqc' + columns <- c( + "Sample", + "raw_total_sequences", + "reads_mapped_percent", + "reads_duplicated_percent" + ) + if (all(columns %in% colnames(mqc_stats))) { + mqc_version <- 'new_mqc' + } else { + stop("Could not find a suitable multiqc table; please provide a correct multiqc.zip file or omit the parameter --multiqc altogether.") + } } else { - stop("Could not find a suitable multiqc table; please provide a correct multiqc.zip file or omit the parameter altogether") + stop("Could not find a suitable multiqc table; please provide a correct multiqc.zip file or omit the parameter --multiqc altogether.") } cat(paste0("*** @@ -911,16 +945,6 @@ sum_NA <- function(x) {if (all(is.na(x))) x[NA_integer_] else sum(x, na.rm = TRU avg_NA <- function(x) {if (all(is.na(x))) x[NA_integer_] else mean(x, na.rm = TRUE)} # Function to average across rows if available, else NA if (mqc_version == 'old_mqc') { - columns <- c( - "Sample", - "FastQC_mqc.generalstats.fastqc.total_sequences", - "FastQC_mqc.generalstats.fastqc.percent_duplicates", - "FastQC_mqc.generalstats.fastqc.percent_gc", - "Cutadapt_mqc.generalstats.cutadapt.percent_trimmed", - "STAR_mqc.generalstats.star.uniquely_mapped_percent", - "featureCounts_mqc.generalstats.featurecounts.percent_assigned" - ) - table_complete <- mqc_stats[,columns] %>% group_by(Sample) %>% dplyr::summarize( @@ -928,13 +952,6 @@ if (mqc_version == 'old_mqc') { across(everything(), avg_NA) ) } else { - columns <- c( - "Sample", - "raw_total_sequences", - "reads_mapped_percent", - "reads_duplicated_percent" - ) - table_complete <- mqc_stats[,columns] %>% group_by(Sample) %>% dplyr::summarize( From 006c69581be4b7f6daba14a1bc4c2b60af232579 Mon Sep 17 00:00:00 2001 From: WackerO Date: Tue, 28 May 2024 15:31:12 +0200 Subject: [PATCH 60/73] Update changelog --- CHANGELOG.md | 1 + 1 file changed, 1 insertion(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index 3491afdd..019af784 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -18,6 +18,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### Fixed +- [#236](https://github.com/qbic-pipelines/rnadeseq/pull/236) Fixed new multiqc check (in case both the files of the old and new mqc version are present) - [#228](https://github.com/qbic-pipelines/rnadeseq/pull/228) Fixed text in report - [#229](https://github.com/qbic-pipelines/rnadeseq/pull/229) Fixed cutoff enrichment plot labels, fixed wrong plotMA function being called (also fixed this changelog) - [#225](https://github.com/qbic-pipelines/rnadeseq/pull/225) Fixed too many devices error from tryCatch around normalized heatmaps From 43ff5ae63069865d32a272503709625db34944a8 Mon Sep 17 00:00:00 2001 From: WackerO Date: Fri, 7 Jun 2024 17:21:03 +0200 Subject: [PATCH 61/73] Working on some more changes that need to be included in the upcoming release --- CHANGELOG.md | 2 +- assets/RNAseq_report.Rmd | 50 ++++++++++++++++++++++++++-------------- 2 files changed, 34 insertions(+), 18 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 3491afdd..4b8ded19 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -3,7 +3,7 @@ The format is based on [Keep a Changelog](https://keepachangelog.com/en/1.0.0/) and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0.html). -## 2.3dev +## 2.3 ### Added diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index 8d79f7f8..7ec8cd1a 100644 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -1305,9 +1305,9 @@ config(pca2, modeBarButtonsToAdd = list(svg_exp)) %>% layout() A sample distance heatmap was generated from the distances of the normalized expression values for all samples. -This visualizes how similar the samples are to each other according to gene expression. +This visualizes how similar the samples are to each other according to the Euclidean distance of gene expression. Two samples have more similar gene expression if their distance is closer to 0 (darker blue). -The original plot can be downloaded [here](./differential_gene_expression/plots/Heatmaps_of_distances.pdf). +The original plot can be downloaded [here](./differential_gene_expression/plots/Heatmaps_of_distances_euclidean.pdf). ```{r heatmap_calculate, include=FALSE} ################## SAMPLE DISTANCES HEATMAP ################## @@ -1317,15 +1317,16 @@ sampleDistClust <- hclust(sampleDists) # Do clustering as input for heatmaps so sampleDistMatrix <- as.matrix(sampleDists) colors = colorRampPalette(rev(RColorBrewer::brewer.pal(9, "Blues")))(255) par(oma=c(3,3,3,3)) +write.table(sampleDistMatrix, file="differential_gene_expression/plots/Heatmaps_of_distances_euclidean.tsv", sep="\t", quote=F) dist_heatmap <- pheatmap(mat = sampleDistMatrix, color=colors, cluster_rows = sampleDistClust, cluster_cols = sampleDistClust) -svg("differential_gene_expression/plots/Heatmaps_of_distances.svg") +svg("differential_gene_expression/plots/Heatmaps_of_distances_euclidean.svg") dist_heatmap dev.off() -png("differential_gene_expression/plots/Heatmaps_of_distances.png") +png("differential_gene_expression/plots/Heatmaps_of_distances_euclidean.png") dist_heatmap dev.off() -pdf("differential_gene_expression/plots/Heatmaps_of_distances.pdf") +pdf("differential_gene_expression/plots/Heatmaps_of_distances_euclidean.pdf") dist_heatmap dev.off() @@ -1367,7 +1368,7 @@ After unzipping the results ZIP file, the differential expression results can be - `DE_genes_tables/`: folder containing one tab-separated table for each of the contrasts in the analysis. Each table contains a list of all differentially expressed genes in the contrast, specifying the mean gene expression across all samples (baseMean), and the log2 fold change value and p-adjusted values (padj) for this contrast. - `final_gene_table/final_gene_list_DESeq2.tsv`: table containing a list of all genes considered in the analysis. Here a summary of the log2 Fold Change and p-adjusted values for all contrasts is displayed. Additionally, the column **filter** shows if this gene was differentially expressed (DE) in any of the contrasts, or not (not_DE). The column **contrast_vector** contains for each contrast considered in the analysis a 1 if the gene was differentially expressed for this contrast or a 0 if it was not. - `plots/`: - - `Heatmaps_of_distances.pdf/.svg`: heatmap of the pairwise euclidean distances among samples, when the `r norm_method_text` normalized gene counts are considered. + - `Heatmaps_of_distances_euclidean.pdf/.svg`: heatmap of the pairwise euclidean distances among samples, when the `r norm_method_text` normalized gene counts are considered. - `PCA_plot.pdf`: PCA of the `r norm_method_text` normalized gene counts. - `boxplots_example_genes/`: boxplots of the normalized gene counts for each of the sample groups for example genes. - `boxplots_requested_genes/`: boxplots of the normalized gene counts for each of the sample groups for the list of requested genes. @@ -1725,8 +1726,7 @@ log2FoldChange_quotient <- length(seq(log2FoldChange_min, log2FoldChange_max, 10 pg <- ggplot(DE_all, aes(x=log2FoldChange, y=logpval, text=paste("Gene: ", gene_name, "
    ", "Log2FC: ", formatC(log2FoldChange, digits=2), "
    ", "-Log10padj: ", formatC(truelogpval, digits=2)))) + - #this goes after alpha=0.5: - geom_jitter(alpha=0.5, width = 0.2, aes(color=ifelse(DE_all$color == "red", "Differentially expressed genes", "Non-differentially expressed genes"))) + + geom_point(alpha=0.5, width = 0.2, aes(color=ifelse(DE_all$color == "red", "Differentially expressed genes", "Non-differentially expressed genes"))) + geom_hline(yintercept = 16, linetype= "dashed", size = 0.2, color = "grey") + geom_hline(yintercept = -log10(params$adj_pval_threshold), size = 0.2, color = "grey") + geom_vline(xintercept = -params$logFC_threshold, size = 0.2, color = "grey") + @@ -2166,10 +2166,12 @@ for (file in contrast_files){ print("------------------------------------") if (nrow(pathway_gostres) > 0){ #if there are enriched pathways - +capture.output(pathway_gostres, file=paste0("pathway_analysis", "/", fname, "/pathway_gostres.tsv")) # Splitting results according to pathway resources (KEGG / REACTOME / GO) res <- split(pathway_gostres, pathway_gostres$source) for (df in res){ + capture.output(df, file=paste0("pathway_analysis", "/", fname, "/df.tsv")) + db_source <- df$source[1] df$short_name <- sapply(df$term_name, substr, start=1, stop=50) @@ -2204,15 +2206,21 @@ for (file in contrast_files){ pathway <- df[i,] gene_list <- unlist(strsplit(pathway$intersection, ",")) mat <- norm_counts[which(norm_counts$Ensembl_ID %in% gene_list), ] - rownames(mat) <- mat$gene_name - mat$gene_name <- NULL - mat$Ensembl_ID <- NULL - mat <- data.matrix(mat) # Skip the heatmaps for CI tests because especially png() works only very unreliably for the heatmaps for some reason if (!isProvided(params$citest) & nrow(mat)>1){ dir.create(paste("pathway_analysis", fname, "pathway_heatmaps", sep="/")) + # Save matrix as table so that the values are accessible if heatmap plotting does not work properly + write.table(mat, file=paste0("pathway_analysis", "/",fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", make.names(pathway$term_id), ".tsv"), sep="\t", quote=F, row.names=F) + write.table(norm_counts, file=paste0("pathway_analysis", "/",fname, "/", "pathway_heatmaps", "/", "norm_counts", pathway$source, "_", make.names(pathway$term_id), ".tsv"), sep="\t", quote=F, row.names=F) + + # Reformat matrix so that it can be used by pheatmap (throws error if String columns/IDs are in the table) + rownames(mat) <- mat$gene_name + mat$gene_name <- NULL + mat$Ensembl_ID <- NULL + mat <- data.matrix(mat) + # Because the image devices sometimes don't work here, surround them with a tryCatch so that the code continues even if a certain heatmap is not saved tryCatch( { @@ -2312,12 +2320,20 @@ if (isProvided(params$path_genelist)){ rownames(norm_counts) <- norm_counts$Ensembl_ID mat <- norm_counts[which(norm_counts$Ensembl_ID %in% gene_list), ] - rownames(mat) <- mat$gene_name - mat$gene_name <- NULL - mat$Ensembl_ID <- NULL - mat <- data.matrix(mat) + write.table(norm_counts, file=paste0(genelist_heatmaps_dir, "/", "norm_counts.tsv"), sep="\t", quote=F, row.names=T) if (nrow(mat)>1){ + write.table(mat, file=paste0(genelist_heatmaps_dir, "/", "mat.tsv"), sep="\t", quote=F, row.names=F) + + # Reformat matrix so that it can be used by pheatmap (throws error if String columns/IDs are in the table) + rownames(mat) <- mat$gene_name + mat$gene_name <- NULL + mat$Ensembl_ID <- NULL + mat <- data.matrix(mat) + + # Save matrix as table so that the values are accessible if heatmap plotting does not work properly + write.table(mat, file=paste0(genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.tsv"), sep="\t", quote=F, row.names=F) + png(filename = paste0(genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.png"), width = 100+ncol(mat)*50, height = 50+nrow(mat)*100) pheatmap(mat = mat, annotation_col = metadata_save, main = "Genelist-specific heatmap of normalized expression values for genes", scale = "row", cluster_cols = params$heatmaps_cluster_cols, cluster_rows = params$heatmaps_cluster_rows) dev.off() From e4e53bbde3e94a3bb22fa32098dd30015ad19118 Mon Sep 17 00:00:00 2001 From: WackerO Date: Mon, 10 Jun 2024 10:42:17 +0200 Subject: [PATCH 62/73] All changes should work now; updated changelog --- CHANGELOG.md | 2 ++ assets/RNAseq_report.Rmd | 34 +++++++++++++++------------------- 2 files changed, 17 insertions(+), 19 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 4b8ded19..bf024826 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -7,6 +7,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### Added +- [#236](https://github.com/qbic-pipelines/rnadeseq/pull/236) Added Euclidian distance/z-score info to report text/heatmaps; added tables next to heatmaps in case the heatmap plotting does not work - [#229](https://github.com/qbic-pipelines/rnadeseq/pull/229) Added param for clustering (or not) the heatmaps - [#226](https://github.com/qbic-pipelines/rnadeseq/pull/226) Added logic to read newer multiqc files from rnaseq 3.12 - [#225](https://github.com/qbic-pipelines/rnadeseq/pull/225) Added param for pathway analysis datasources @@ -14,6 +15,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### Changed +- [#236](https://github.com/qbic-pipelines/rnadeseq/pull/236) Changed geom_jitter to geom_point for volcano plot so that points are accurate - [#225](https://github.com/qbic-pipelines/rnadeseq/pull/225) Shortened names of some output files ### Fixed diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index 7ec8cd1a..636fd678 100644 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -1303,11 +1303,11 @@ config(pca2, modeBarButtonsToAdd = list(svg_exp)) %>% layout() ## Sample distance heatmap -A sample distance heatmap was generated from the distances of the normalized expression values for all samples. +A sample distance heatmap was generated from the Euclidean distances of the normalized expression values for all samples. -This visualizes how similar the samples are to each other according to the Euclidean distance of gene expression. +This visualizes how similar the samples are to each other according to gene expression. Two samples have more similar gene expression if their distance is closer to 0 (darker blue). -The original plot can be downloaded [here](./differential_gene_expression/plots/Heatmaps_of_distances_euclidean.pdf). +The original plot can be downloaded [here](./differential_gene_expression/plots/Heatmaps_of_distances.pdf). ```{r heatmap_calculate, include=FALSE} ################## SAMPLE DISTANCES HEATMAP ################## @@ -1317,16 +1317,16 @@ sampleDistClust <- hclust(sampleDists) # Do clustering as input for heatmaps so sampleDistMatrix <- as.matrix(sampleDists) colors = colorRampPalette(rev(RColorBrewer::brewer.pal(9, "Blues")))(255) par(oma=c(3,3,3,3)) -write.table(sampleDistMatrix, file="differential_gene_expression/plots/Heatmaps_of_distances_euclidean.tsv", sep="\t", quote=F) +write.table(sampleDistMatrix, file="differential_gene_expression/plots/Heatmaps_of_distances.tsv", sep="\t", quote=F) dist_heatmap <- pheatmap(mat = sampleDistMatrix, color=colors, cluster_rows = sampleDistClust, cluster_cols = sampleDistClust) -svg("differential_gene_expression/plots/Heatmaps_of_distances_euclidean.svg") +svg("differential_gene_expression/plots/Heatmaps_of_distances.svg") dist_heatmap dev.off() -png("differential_gene_expression/plots/Heatmaps_of_distances_euclidean.png") +png("differential_gene_expression/plots/Heatmaps_of_distances.png") dist_heatmap dev.off() -pdf("differential_gene_expression/plots/Heatmaps_of_distances_euclidean.pdf") +pdf("differential_gene_expression/plots/Heatmaps_of_distances.pdf") dist_heatmap dev.off() @@ -1368,7 +1368,7 @@ After unzipping the results ZIP file, the differential expression results can be - `DE_genes_tables/`: folder containing one tab-separated table for each of the contrasts in the analysis. Each table contains a list of all differentially expressed genes in the contrast, specifying the mean gene expression across all samples (baseMean), and the log2 fold change value and p-adjusted values (padj) for this contrast. - `final_gene_table/final_gene_list_DESeq2.tsv`: table containing a list of all genes considered in the analysis. Here a summary of the log2 Fold Change and p-adjusted values for all contrasts is displayed. Additionally, the column **filter** shows if this gene was differentially expressed (DE) in any of the contrasts, or not (not_DE). The column **contrast_vector** contains for each contrast considered in the analysis a 1 if the gene was differentially expressed for this contrast or a 0 if it was not. - `plots/`: - - `Heatmaps_of_distances_euclidean.pdf/.svg`: heatmap of the pairwise euclidean distances among samples, when the `r norm_method_text` normalized gene counts are considered. + - `Heatmaps_of_distances.pdf/.png/.svg`: heatmap of the pairwise euclidean distances among samples, when the `r norm_method_text` normalized gene counts are considered. - `PCA_plot.pdf`: PCA of the `r norm_method_text` normalized gene counts. - `boxplots_example_genes/`: boxplots of the normalized gene counts for each of the sample groups for example genes. - `boxplots_requested_genes/`: boxplots of the normalized gene counts for each of the sample groups for the list of requested genes. @@ -2166,11 +2166,10 @@ for (file in contrast_files){ print("------------------------------------") if (nrow(pathway_gostres) > 0){ #if there are enriched pathways -capture.output(pathway_gostres, file=paste0("pathway_analysis", "/", fname, "/pathway_gostres.tsv")) + # Splitting results according to pathway resources (KEGG / REACTOME / GO) res <- split(pathway_gostres, pathway_gostres$source) for (df in res){ - capture.output(df, file=paste0("pathway_analysis", "/", fname, "/df.tsv")) db_source <- df$source[1] df$short_name <- sapply(df$term_name, substr, start=1, stop=50) @@ -2213,7 +2212,6 @@ capture.output(pathway_gostres, file=paste0("pathway_analysis", "/", fname, "/pa # Save matrix as table so that the values are accessible if heatmap plotting does not work properly write.table(mat, file=paste0("pathway_analysis", "/",fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", make.names(pathway$term_id), ".tsv"), sep="\t", quote=F, row.names=F) - write.table(norm_counts, file=paste0("pathway_analysis", "/",fname, "/", "pathway_heatmaps", "/", "norm_counts", pathway$source, "_", make.names(pathway$term_id), ".tsv"), sep="\t", quote=F, row.names=F) # Reformat matrix so that it can be used by pheatmap (throws error if String columns/IDs are in the table) rownames(mat) <- mat$gene_name @@ -2225,7 +2223,7 @@ capture.output(pathway_gostres, file=paste0("pathway_analysis", "/", fname, "/pa tryCatch( { png(filename = paste0("pathway_analysis", "/",fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", make.names(pathway$term_id), ".png"), width = ifelse(nrow(mat)<=20, 200, 125)+ncol(mat)*40, height = 50+nrow(mat)*50) - pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = params$heatmaps_cluster_cols, cluster_rows = params$heatmaps_cluster_rows) + pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values (z-scores) for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = params$heatmaps_cluster_cols, cluster_rows = params$heatmaps_cluster_rows) dev.off() }, error=function(e) { @@ -2235,7 +2233,7 @@ capture.output(pathway_gostres, file=paste0("pathway_analysis", "/", fname, "/pa tryCatch( { pdf(paste0("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", make.names(pathway$term_id), ".pdf"), width = ifelse(nrow(mat)<=20, 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005) - pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = params$heatmaps_cluster_cols, cluster_rows = params$heatmaps_cluster_rows) + pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values (z-scores) for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = params$heatmaps_cluster_cols, cluster_rows = params$heatmaps_cluster_rows) dev.off() }, error=function(e) { @@ -2245,7 +2243,7 @@ capture.output(pathway_gostres, file=paste0("pathway_analysis", "/", fname, "/pa tryCatch( { svg(paste0("pathway_analysis", "/", fname, "/", "pathway_heatmaps", "/", "Heatmap_normalized_counts_", pathway$source, "_", make.names(pathway$term_id), ".svg"), width = ifelse(nrow(mat)<=20, 50, 10)+ncol(mat)*0.1, height = 5+nrow(mat)*0.005) - pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = params$heatmaps_cluster_cols, cluster_rows = params$heatmaps_cluster_rows) + pheatmap(mat = mat, annotation_col = metadata_save, main = paste("Heatmap of normalized expression values (z-scores) for genes in pathway ", pathway$short_name, "(",pathway$source,")",sep=" "), scale = "row", cluster_cols = params$heatmaps_cluster_cols, cluster_rows = params$heatmaps_cluster_rows) dev.off() }, error=function(e) { @@ -2320,10 +2318,8 @@ if (isProvided(params$path_genelist)){ rownames(norm_counts) <- norm_counts$Ensembl_ID mat <- norm_counts[which(norm_counts$Ensembl_ID %in% gene_list), ] - write.table(norm_counts, file=paste0(genelist_heatmaps_dir, "/", "norm_counts.tsv"), sep="\t", quote=F, row.names=T) if (nrow(mat)>1){ - write.table(mat, file=paste0(genelist_heatmaps_dir, "/", "mat.tsv"), sep="\t", quote=F, row.names=F) # Reformat matrix so that it can be used by pheatmap (throws error if String columns/IDs are in the table) rownames(mat) <- mat$gene_name @@ -2335,13 +2331,13 @@ if (isProvided(params$path_genelist)){ write.table(mat, file=paste0(genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.tsv"), sep="\t", quote=F, row.names=F) png(filename = paste0(genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.png"), width = 100+ncol(mat)*50, height = 50+nrow(mat)*100) - pheatmap(mat = mat, annotation_col = metadata_save, main = "Genelist-specific heatmap of normalized expression values for genes", scale = "row", cluster_cols = params$heatmaps_cluster_cols, cluster_rows = params$heatmaps_cluster_rows) + pheatmap(mat = mat, annotation_col = metadata_save, main = "Genelist-specific Heatmap of normalized expression values (z-scores) for genes", scale = "row", cluster_cols = params$heatmaps_cluster_cols, cluster_rows = params$heatmaps_cluster_rows) dev.off() pdf(paste0(genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.pdf"), width = 10+ncol(mat)*0.05, height = 5+nrow(mat)*0.005) - pheatmap(mat = mat, annotation_col = metadata_save, main = "Genelist-specific heatmap of normalized expression values for genes", scale = "row", cluster_cols = params$heatmaps_cluster_cols, cluster_rows = params$heatmaps_cluster_rows) + pheatmap(mat = mat, annotation_col = metadata_save, main = "Genelist-specific Heatmap of normalized expression values (z-scores) for genes", scale = "row", cluster_cols = params$heatmaps_cluster_cols, cluster_rows = params$heatmaps_cluster_rows) dev.off() svg(paste0(genelist_heatmaps_dir, "/", "Heatmap_normalized_counts_gene_list.svg"), width = 10+ncol(mat)*0.1, height = 5+nrow(mat)*0.005) - pheatmap(mat = mat, annotation_col = metadata_save, main = "Genelist-specific heatmap of normalized expression values for genes", scale = "row", cluster_cols = params$heatmaps_cluster_cols, cluster_rows = params$heatmaps_cluster_rows) + pheatmap(mat = mat, annotation_col = metadata_save, main = "Genelist-specific Heatmap of normalized expression values (z-scores) for genes", scale = "row", cluster_cols = params$heatmaps_cluster_cols, cluster_rows = params$heatmaps_cluster_rows) dev.off() } } From 0565bb475712af6b2c243235dff207d6e7b88105 Mon Sep 17 00:00:00 2001 From: WackerO Date: Mon, 10 Jun 2024 10:45:30 +0200 Subject: [PATCH 63/73] fix changelog --- CHANGELOG.md | 5 +++-- 1 file changed, 3 insertions(+), 2 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index bf024826..25d440ed 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -7,7 +7,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### Added -- [#236](https://github.com/qbic-pipelines/rnadeseq/pull/236) Added Euclidian distance/z-score info to report text/heatmaps; added tables next to heatmaps in case the heatmap plotting does not work +- [#237](https://github.com/qbic-pipelines/rnadeseq/pull/237) Added Euclidian distance/z-score info to report text/heatmaps; added tables next to heatmaps in case the heatmap plotting does not work - [#229](https://github.com/qbic-pipelines/rnadeseq/pull/229) Added param for clustering (or not) the heatmaps - [#226](https://github.com/qbic-pipelines/rnadeseq/pull/226) Added logic to read newer multiqc files from rnaseq 3.12 - [#225](https://github.com/qbic-pipelines/rnadeseq/pull/225) Added param for pathway analysis datasources @@ -15,11 +15,12 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### Changed -- [#236](https://github.com/qbic-pipelines/rnadeseq/pull/236) Changed geom_jitter to geom_point for volcano plot so that points are accurate +- [#237](https://github.com/qbic-pipelines/rnadeseq/pull/237) Changed geom_jitter to geom_point for volcano plot so that points are accurate - [#225](https://github.com/qbic-pipelines/rnadeseq/pull/225) Shortened names of some output files ### Fixed +- [#236](https://github.com/qbic-pipelines/rnadeseq/pull/236) Fix new multiqc (add logic to better check which multiqc version is provided (sometimes both the old and the new table file are present in the mqc)) - [#228](https://github.com/qbic-pipelines/rnadeseq/pull/228) Fixed text in report - [#229](https://github.com/qbic-pipelines/rnadeseq/pull/229) Fixed cutoff enrichment plot labels, fixed wrong plotMA function being called (also fixed this changelog) - [#225](https://github.com/qbic-pipelines/rnadeseq/pull/225) Fixed too many devices error from tryCatch around normalized heatmaps From 072623429da09309f66721bc9a73247c39cb7dce Mon Sep 17 00:00:00 2001 From: WackerO Date: Mon, 10 Jun 2024 11:14:08 +0200 Subject: [PATCH 64/73] minor fixes for release --- CHANGELOG.md | 1 + conf/test_custom_gmt.config | 1 - docs/usage.md | 4 ++-- 3 files changed, 3 insertions(+), 3 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 25d440ed..4632f2af 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -20,6 +20,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### Fixed +- [#237](https://github.com/qbic-pipelines/rnadeseq/pull/237) Fix version in usage docs and remove duplicate datasources in test_custom_gmt.config - [#236](https://github.com/qbic-pipelines/rnadeseq/pull/236) Fix new multiqc (add logic to better check which multiqc version is provided (sometimes both the old and the new table file are present in the mqc)) - [#228](https://github.com/qbic-pipelines/rnadeseq/pull/228) Fixed text in report - [#229](https://github.com/qbic-pipelines/rnadeseq/pull/229) Fixed cutoff enrichment plot labels, fixed wrong plotMA function being called (also fixed this changelog) diff --git a/conf/test_custom_gmt.config b/conf/test_custom_gmt.config index 1e901589..3ee2eab9 100644 --- a/conf/test_custom_gmt.config +++ b/conf/test_custom_gmt.config @@ -31,5 +31,4 @@ params { datasources = 'KEGG,REAC' genome = 'GRCm38' custom_gmt = 'https://raw.githubusercontent.com/qbic-pipelines/rnadeseq/dev/testdata/gprofiler_full_mmusculus.ENSG.gmt' - datasources = 'KEGG,REAC' } diff --git a/docs/usage.md b/docs/usage.md index ba2dac7c..05da6ed0 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -89,7 +89,7 @@ The `qbic-pipelines/rnadeseq` pipeline relies on the output from the `nf-core/rn The typical command for running the pipeline is as follows: ```bash -nextflow run qbic-pipelines/rnadeseq -r 2.2 -profile docker \ +nextflow run qbic-pipelines/rnadeseq -r 2.3 -profile docker \ --gene_counts 'merged_gene_counts.txt' \ --input 'QXXXX_sample_preparations.tsv' \ --model 'linear_model.txt' \ @@ -157,7 +157,7 @@ curl https://codeload.github.com/qbic-pipelines/rnadeseq/tar.gz/master | tar -xz Afterwards, you should be able to also run test_star_rsem and test_star_salmon from qbic-pipelines/rnadeseq without manually cloning, e.g.: ```bash -nextflow run qbic-pipelines/rnadeseq -r 2.2 -profile docker,test_star_salmon +nextflow run qbic-pipelines/rnadeseq -r 2.3 -profile docker,test_star_salmon ``` ### Reproducibility From f52a95e8ea43aa93bbe77aa2623c5ef85714072b Mon Sep 17 00:00:00 2001 From: WackerO <43847497+WackerO@users.noreply.github.com> Date: Mon, 10 Jun 2024 14:55:43 +0200 Subject: [PATCH 65/73] Update assets/RNAseq_report.Rmd Co-authored-by: Till E. <64961761+tillenglert@users.noreply.github.com> --- assets/RNAseq_report.Rmd | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/assets/RNAseq_report.Rmd b/assets/RNAseq_report.Rmd index e820b124..9e66e7d4 100644 --- a/assets/RNAseq_report.Rmd +++ b/assets/RNAseq_report.Rmd @@ -1385,7 +1385,7 @@ After unzipping the results ZIP file, the differential expression results can be - `DE_genes_tables/`: folder containing one tab-separated table for each of the contrasts in the analysis. Each table contains a list of all differentially expressed genes in the contrast, specifying the mean gene expression across all samples (baseMean), and the log2 fold change value and p-adjusted values (padj) for this contrast. - `final_gene_table/final_gene_list_DESeq2.tsv`: table containing a list of all genes considered in the analysis. Here a summary of the log2 Fold Change and p-adjusted values for all contrasts is displayed. Additionally, the column **filter** shows if this gene was differentially expressed (DE) in any of the contrasts, or not (not_DE). The column **contrast_vector** contains for each contrast considered in the analysis a 1 if the gene was differentially expressed for this contrast or a 0 if it was not. - `plots/`: - - `Heatmaps_of_distances.pdf/.png/.svg`: heatmap of the pairwise euclidean distances among samples, when the `r norm_method_text` normalized gene counts are considered. + - `Heatmaps_of_distances.pdf/.png/.svg`: heatmap of the pairwise Euclidean distances among samples, when the `r norm_method_text` normalized gene counts are considered. - `PCA_plot.pdf`: PCA of the `r norm_method_text` normalized gene counts. - `boxplots_example_genes/`: boxplots of the normalized gene counts for each of the sample groups for example genes. - `boxplots_requested_genes/`: boxplots of the normalized gene counts for each of the sample groups for the list of requested genes. From a915bdb181f43d0c237a2694d44e8fe0dad51321 Mon Sep 17 00:00:00 2001 From: WackerO <43847497+WackerO@users.noreply.github.com> Date: Wed, 12 Jun 2024 14:29:48 +0200 Subject: [PATCH 66/73] Update assets/methods_description_template.yml Co-authored-by: Daniel Straub <42973691+d4straub@users.noreply.github.com> --- assets/methods_description_template.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/assets/methods_description_template.yml b/assets/methods_description_template.yml index f5f5bfe4..b0f33b73 100644 --- a/assets/methods_description_template.yml +++ b/assets/methods_description_template.yml @@ -6,7 +6,7 @@ plot_type: "html" ## You inject any metadata in the Nextflow '${workflow}' object data: |

    Methods

    -

    Data was processed using qbic-pipelines/rnadeseq v${workflow.manifest.version} ${doi_text} of the nf-core collection of workflows (Ewels et al., 2020), utilising reproducible software environments from the Bioconda (Grüning et al., 2018) and Biocontainers (da Veiga Leprevost et al., 2017) projects.

    +

    Data was processed using qbic-pipelines/rnadeseq v${workflow.manifest.version} ${doi_text} adapted from the nf-core workflow template (Ewels et al., 2020), utilising reproducible software environments from the Bioconda (Grüning et al., 2018) and Biocontainers (da Veiga Leprevost et al., 2017) projects.

    The pipeline was executed with Nextflow v${workflow.nextflow.version} (Di Tommaso et al., 2017) with the following command:

    ${workflow.commandLine}

    ${tool_citations}

    From 428df02e23238d81f434f7eb5c758d095fe21724 Mon Sep 17 00:00:00 2001 From: WackerO Date: Wed, 12 Jun 2024 14:47:56 +0200 Subject: [PATCH 67/73] Fix branch protection bug --- .github/workflows/branch.yml | 2 +- CHANGELOG.md | 1 + 2 files changed, 2 insertions(+), 1 deletion(-) diff --git a/.github/workflows/branch.yml b/.github/workflows/branch.yml index fd4d331d..d64b4c99 100644 --- a/.github/workflows/branch.yml +++ b/.github/workflows/branch.yml @@ -13,7 +13,7 @@ jobs: - name: Check PRs if: github.repository == 'qbic-pipelines/rnadeseq' run: | - "{ [[ ${{github.event.pull_request.head.repo.full_name }} == qbic-pipelines/rnadeseq ]] && [[ $GITHUB_HEAD_REF = "dev" ]]; } || [[ $GITHUB_HEAD_REF == "patch" ]]" + { [[ ${{github.event.pull_request.head.repo.full_name }} == qbic-pipelines/rnadeseq ]] && [[ $GITHUB_HEAD_REF == "dev" ]]; } || [[ $GITHUB_HEAD_REF == "patch" ]] # If the above check failed, post a comment on the PR explaining the failure # NOTE - this doesn't currently work if the PR is coming from a fork, due to limitations in GitHub actions secrets diff --git a/CHANGELOG.md b/CHANGELOG.md index 81393416..26e1f80f 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -20,6 +20,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### Fixed +- [#238](https://github.com/qbic-pipelines/rnadeseq/pull/238) Fix branch protection bug - [#237](https://github.com/qbic-pipelines/rnadeseq/pull/237) Fix version in usage docs and remove duplicate datasources in test_custom_gmt.config - [#236](https://github.com/qbic-pipelines/rnadeseq/pull/236) Fixed new multiqc check (in case both the files of the old and new mqc version are present) - [#234](https://github.com/qbic-pipelines/rnadeseq/pull/234) Fixed unnecessary file permission changes from #225 From 5dc13c851db0c1dc50d082509728431e62ffab9e Mon Sep 17 00:00:00 2001 From: WackerO Date: Wed, 12 Jun 2024 15:18:17 +0200 Subject: [PATCH 68/73] second attempt at fixing branch protection --- .github/workflows/branch.yml | 8 ++++---- CHANGELOG.md | 1 + 2 files changed, 5 insertions(+), 4 deletions(-) diff --git a/.github/workflows/branch.yml b/.github/workflows/branch.yml index d64b4c99..be960188 100644 --- a/.github/workflows/branch.yml +++ b/.github/workflows/branch.yml @@ -11,11 +11,11 @@ jobs: steps: # PRs to the nf-core repo master branch are only ok if coming from the nf-core repo `dev` or any `patch` branches - name: Check PRs - if: github.repository == 'qbic-pipelines/rnadeseq' + if: github.repository == '{{ name }}' run: | - { [[ ${{github.event.pull_request.head.repo.full_name }} == qbic-pipelines/rnadeseq ]] && [[ $GITHUB_HEAD_REF == "dev" ]]; } || [[ $GITHUB_HEAD_REF == "patch" ]] + { [[ {% raw %}${{github.event.pull_request.head.repo.full_name }}{% endraw %} == {{ name }} ]] && [[ $GITHUB_HEAD_REF == "dev" ]]; } || [[ $GITHUB_HEAD_REF == "patch" ]] - # If the above check failed, post a comment on the PR explaining the failure + # If the above check failed, post a comment on the PR explaining the failure {%- raw %} # NOTE - this doesn't currently work if the PR is coming from a fork, due to limitations in GitHub actions secrets - name: Post PR comment if: failure() @@ -41,4 +41,4 @@ jobs: Thanks again for your contribution! repo-token: ${{ secrets.GITHUB_TOKEN }} - allow-repeats: false + allow-repeats: false {%- endraw %} \ No newline at end of file diff --git a/CHANGELOG.md b/CHANGELOG.md index 26e1f80f..ed85b190 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -20,6 +20,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### Fixed +- [#239](https://github.com/qbic-pipelines/rnadeseq/pull/239) Fix branch protection bug --> second attempt - [#238](https://github.com/qbic-pipelines/rnadeseq/pull/238) Fix branch protection bug - [#237](https://github.com/qbic-pipelines/rnadeseq/pull/237) Fix version in usage docs and remove duplicate datasources in test_custom_gmt.config - [#236](https://github.com/qbic-pipelines/rnadeseq/pull/236) Fixed new multiqc check (in case both the files of the old and new mqc version are present) From 1b765e07da62be409e68343b5a2ad928a4638d0d Mon Sep 17 00:00:00 2001 From: WackerO Date: Wed, 12 Jun 2024 15:26:18 +0200 Subject: [PATCH 69/73] third attempt at fixing branch protection --- .github/workflows/branch.yml | 4 ++-- CHANGELOG.md | 1 + 2 files changed, 3 insertions(+), 2 deletions(-) diff --git a/.github/workflows/branch.yml b/.github/workflows/branch.yml index be960188..d842235a 100644 --- a/.github/workflows/branch.yml +++ b/.github/workflows/branch.yml @@ -11,9 +11,9 @@ jobs: steps: # PRs to the nf-core repo master branch are only ok if coming from the nf-core repo `dev` or any `patch` branches - name: Check PRs - if: github.repository == '{{ name }}' + if: github.repository == 'qbic-pipelines/rnadeseq' run: | - { [[ {% raw %}${{github.event.pull_request.head.repo.full_name }}{% endraw %} == {{ name }} ]] && [[ $GITHUB_HEAD_REF == "dev" ]]; } || [[ $GITHUB_HEAD_REF == "patch" ]] + { [[ {% raw %}${{github.event.pull_request.head.repo.full_name }}{% endraw %} == qbic-pipelines/rnadeseq ]] && [[ $GITHUB_HEAD_REF == "dev" ]]; } || [[ $GITHUB_HEAD_REF == "patch" ]] # If the above check failed, post a comment on the PR explaining the failure {%- raw %} # NOTE - this doesn't currently work if the PR is coming from a fork, due to limitations in GitHub actions secrets diff --git a/CHANGELOG.md b/CHANGELOG.md index ed85b190..89adac52 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -20,6 +20,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### Fixed +- [#240](https://github.com/qbic-pipelines/rnadeseq/pull/240) Fix branch protection bug --> third attempt - [#239](https://github.com/qbic-pipelines/rnadeseq/pull/239) Fix branch protection bug --> second attempt - [#238](https://github.com/qbic-pipelines/rnadeseq/pull/238) Fix branch protection bug - [#237](https://github.com/qbic-pipelines/rnadeseq/pull/237) Fix version in usage docs and remove duplicate datasources in test_custom_gmt.config From 8fd4aca8cc0527121b2d4a76d21a587b9ac531f7 Mon Sep 17 00:00:00 2001 From: WackerO Date: Thu, 13 Jun 2024 08:14:34 +0200 Subject: [PATCH 70/73] undid branch.yml changes --- .github/workflows/branch.yml | 2 +- CHANGELOG.md | 1 + 2 files changed, 2 insertions(+), 1 deletion(-) diff --git a/.github/workflows/branch.yml b/.github/workflows/branch.yml index d842235a..64d571fc 100644 --- a/.github/workflows/branch.yml +++ b/.github/workflows/branch.yml @@ -13,7 +13,7 @@ jobs: - name: Check PRs if: github.repository == 'qbic-pipelines/rnadeseq' run: | - { [[ {% raw %}${{github.event.pull_request.head.repo.full_name }}{% endraw %} == qbic-pipelines/rnadeseq ]] && [[ $GITHUB_HEAD_REF == "dev" ]]; } || [[ $GITHUB_HEAD_REF == "patch" ]] + "{ [[ ${{github.event.pull_request.head.repo.full_name }} == qbic-pipelines/rnadeseq ]] && [[ $GITHUB_HEAD_REF = "dev" ]]; } || [[ $GITHUB_HEAD_REF == "patch" ]]" # If the above check failed, post a comment on the PR explaining the failure {%- raw %} # NOTE - this doesn't currently work if the PR is coming from a fork, due to limitations in GitHub actions secrets diff --git a/CHANGELOG.md b/CHANGELOG.md index 89adac52..039ed3dc 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -20,6 +20,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### Fixed +- [#241](https://github.com/qbic-pipelines/rnadeseq/pull/241) Undo fix branch protection bug in order to get precommit test to run - [#240](https://github.com/qbic-pipelines/rnadeseq/pull/240) Fix branch protection bug --> third attempt - [#239](https://github.com/qbic-pipelines/rnadeseq/pull/239) Fix branch protection bug --> second attempt - [#238](https://github.com/qbic-pipelines/rnadeseq/pull/238) Fix branch protection bug From b579def6764c43c490e332b7e47e42ddba788409 Mon Sep 17 00:00:00 2001 From: WackerO Date: Thu, 13 Jun 2024 08:20:45 +0200 Subject: [PATCH 71/73] prettier --- .github/workflows/branch.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/.github/workflows/branch.yml b/.github/workflows/branch.yml index 64d571fc..c38a1cb0 100644 --- a/.github/workflows/branch.yml +++ b/.github/workflows/branch.yml @@ -41,4 +41,4 @@ jobs: Thanks again for your contribution! repo-token: ${{ secrets.GITHUB_TOKEN }} - allow-repeats: false {%- endraw %} \ No newline at end of file + allow-repeats: false {%- endraw %} From 16576921aee491b9a398ff387bd899ea7432705c Mon Sep 17 00:00:00 2001 From: WackerO Date: Thu, 13 Jun 2024 11:32:15 +0200 Subject: [PATCH 72/73] fixed path issue with report file --- modules/local/report.nf | 4 +++- nextflow.config | 2 ++ nextflow_schema.json | 10 ++++++++++ workflows/rnadeseq.nf | 6 +++++- 4 files changed, 20 insertions(+), 2 deletions(-) diff --git a/modules/local/report.nf b/modules/local/report.nf index 413e241c..5c0fcab1 100644 --- a/modules/local/report.nf +++ b/modules/local/report.nf @@ -19,6 +19,8 @@ process REPORT { path multiqc path custom_gmt path custom_background + path report_file + path references_file output: path "*.zip" @@ -54,7 +56,7 @@ process REPORT { mv MultiQC/multiqc_plots/ MultiQC/multiqc_data/ MultiQC/multiqc_report.html QC/ || mv multiqc/*/multiqc_plots/ multiqc/*/multiqc_data/ multiqc/*/multiqc_report.html QC/ || mv multiqc_plots/ multiqc_data/ multiqc_report.html QC/ fi Execute_report.R \ - --report '$baseDir/assets/RNAseq_report.Rmd' \ + --report '$report_file' \ --output 'RNAseq_report.html' \ --input_type $params.input_type \ --gene_counts $gene_counts \ diff --git a/nextflow.config b/nextflow.config index fe2d3897..8db8337c 100644 --- a/nextflow.config +++ b/nextflow.config @@ -48,6 +48,8 @@ params { multiqc = 'NO_FILE4' quote = 'NO_FILE5' software_versions = 'NO_FILE6' + report_file = "$projectDir/assets/RNAseq_report.Rmd" + references_file = "$projectDir/assets/references.bibtex" // Additional arg to indicate that a github test is run citest = false diff --git a/nextflow_schema.json b/nextflow_schema.json index b1b923d6..24b665eb 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -183,6 +183,16 @@ "type": "string", "default": "NO_FILE6", "description": "CSV/YML containing nf-core/rnaseq software versions." + }, + "report_file": { + "type": "string", + "default": "$projectDir/assets/RNAseq_report.Rmd", + "description": "Report Rmd file, should not be changed." + }, + "references_file": { + "type": "string", + "default": "$projectDir/assets/references.bibtex", + "description": "Reference bibtex file for the report, should not be changed." } } }, diff --git a/workflows/rnadeseq.nf b/workflows/rnadeseq.nf index a47b3a0c..ead6be2a 100644 --- a/workflows/rnadeseq.nf +++ b/workflows/rnadeseq.nf @@ -31,6 +31,8 @@ ch_custom_gmt = Channel.fromPath(params.custom_gmt) ch_custom_background = Channel.fromPath(params.custom_background) ch_proj_summary_file = Channel.fromPath(params.project_summary) ch_softwareversions_file = Channel.fromPath(params.software_versions) +ch_report_file = Channel.fromPath(params.report_file) +ch_references_file = Channel.fromPath(params.references_file) /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ @@ -74,7 +76,9 @@ workflow RNADESEQ { ch_softwareversions_file, ch_multiqc_file, ch_custom_gmt, - ch_custom_background + ch_custom_background, + ch_report_file, + ch_references_file ) //TODO: Enable this: From 3863f7bf2c662ec2175e2cc34c72cffdbe2860a1 Mon Sep 17 00:00:00 2001 From: WackerO Date: Thu, 13 Jun 2024 12:38:05 +0200 Subject: [PATCH 73/73] added two params to nfcore.yml to suppress error --- .nf-core.yml | 2 ++ nextflow_schema.json | 4 ++-- 2 files changed, 4 insertions(+), 2 deletions(-) diff --git a/.nf-core.yml b/.nf-core.yml index 4ce60765..ee90d841 100644 --- a/.nf-core.yml +++ b/.nf-core.yml @@ -21,6 +21,8 @@ lint: - manifest.homePage - config_defaults: - params.custom_config_base + - params.report_file + - params.references_file multiqc_config: - report_comment # otherwise throws error: multiqc_config: 'assets/multiqc_config.yml' does not contain a matching 'report_comment'. diff --git a/nextflow_schema.json b/nextflow_schema.json index 24b665eb..aa6b89a2 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -186,12 +186,12 @@ }, "report_file": { "type": "string", - "default": "$projectDir/assets/RNAseq_report.Rmd", + "default": "${projectDir}/assets/RNAseq_report.Rmd", "description": "Report Rmd file, should not be changed." }, "references_file": { "type": "string", - "default": "$projectDir/assets/references.bibtex", + "default": "${projectDir}/assets/references.bibtex", "description": "Reference bibtex file for the report, should not be changed." } }

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z$3Si?`1rN&U-%Z=K57&(f=vjiP@OmmOUW+RPOd2{b5|69h%j*@-W}>bot8g;_JCq%DL1L6jNdmJ zzpq;lDd$bKl0GLp)szjMv+UeW;=bsKmz-(OmD4cZ5Em+bha6ALCEi7lda;cY?O8|m zdVhQ-VjfP+HF=P$IeUxzyu+_9bKisXnn9?g(A=vvgDomtXPvZ~G2TxN(#yQaa_Q7C z@LOgUW<3jE2M)S(;iE76u(0uYZR?7yhLruNX1r|$A45*o2DLJT_v(a!ZrX9i#gU2@ zobig5Ty~eq;I?+V=OK*+UYs!VxU>fsq&8Gdx)dy+w-A8flE0@1b$P2U{Tu!;0#*Fq zzw1I5>3{!Dqy5(oe|y1y?eKrGgCrHeWTyVV8;HgysFRximl6KE`zUMu*X#c4GylI2 aEUI}0*A%Yv8~ls146@Virw2Pc&i@a1yY|lj literal 0 HcmV?d00001 diff --git a/assets/samplesheet.csv b/assets/samplesheet.csv new file mode 100644 index 00000000..5f653ab7 --- /dev/null +++ b/assets/samplesheet.csv @@ -0,0 +1,3 @@ +sample,fastq_1,fastq_2 +SAMPLE_PAIRED_END,/path/to/fastq/files/AEG588A1_S1_L002_R1_001.fastq.gz,/path/to/fastq/files/AEG588A1_S1_L002_R2_001.fastq.gz +SAMPLE_SINGLE_END,/path/to/fastq/files/AEG588A4_S4_L003_R1_001.fastq.gz, diff --git a/assets/schema_input.json b/assets/schema_input.json new file mode 100644 index 00000000..43cd7c69 --- /dev/null +++ b/assets/schema_input.json @@ -0,0 +1,33 @@ +{ + "$schema": "http://json-schema.org/draft-07/schema", + "$id": "https://raw.githubusercontent.com/nf-core/rnadeseq/master/assets/schema_input.json", + "title": "nf-core/rnadeseq pipeline - params.input schema", + "description": "Schema for the file provided with params.input", + "type": "array", + "items": { + "type": "object", + "properties": { + "sample": { + "type": "string", + "pattern": "^\\S+$", + "errorMessage": "Sample name must be provided and cannot contain spaces", + "meta": ["id"] + }, + "fastq_1": { + "type": "string", + "format": "file-path", + "exists": true, + "pattern": "^\\S+\\.f(ast)?q\\.gz$", + "errorMessage": "FastQ file for reads 1 must be provided, cannot contain spaces and must have extension '.fq.gz' or '.fastq.gz'" + }, + "fastq_2": { + "type": "string", + "format": "file-path", + "exists": true, + "pattern": "^\\S+\\.f(ast)?q\\.gz$", + "errorMessage": "FastQ file for reads 2 cannot contain spaces and must have extension '.fq.gz' or '.fastq.gz'" + } + }, + "required": ["sample", "fastq_1"] + } +} diff --git a/assets/sendmail_template.txt b/assets/sendmail_template.txt new file mode 100644 index 00000000..953c8470 --- /dev/null +++ b/assets/sendmail_template.txt @@ -0,0 +1,53 @@ +To: $email +Subject: $subject +Mime-Version: 1.0 +Content-Type: multipart/related;boundary="nfcoremimeboundary" + +--nfcoremimeboundary +Content-Type: text/html; charset=utf-8 + +$email_html + +--nfcoremimeboundary +Content-Type: image/png;name="nf-core-rnadeseq_logo.png" +Content-Transfer-Encoding: base64 +Content-ID: +Content-Disposition: inline; filename="nf-core-rnadeseq_logo_light.png" + +<% out << new File("$projectDir/assets/nf-core-rnadeseq_logo_light.png"). + bytes. + encodeBase64(). + toString(). + tokenize( '\n' )*. + toList()*. + collate( 76 )*. + collect { it.join() }. + flatten(). + join( '\n' ) %> + +<% +if (mqcFile){ +def mqcFileObj = new File("$mqcFile") +if (mqcFileObj.length() < mqcMaxSize){ +out << """ +--nfcoremimeboundary +Content-Type: text/html; name=\"multiqc_report\" +Content-Transfer-Encoding: base64 +Content-ID: +Content-Disposition: attachment; filename=\"${mqcFileObj.getName()}\" + +${mqcFileObj. + bytes. + encodeBase64(). + toString(). + tokenize( '\n' )*. + toList()*. + collate( 76 )*. + collect { it.join() }. + flatten(). + join( '\n' )} +""" +}} +%> + +--nfcoremimeboundary-- diff --git a/assets/slackreport.json b/assets/slackreport.json new file mode 100644 index 00000000..a4bb18d3 --- /dev/null +++ b/assets/slackreport.json @@ -0,0 +1,34 @@ +{ + "attachments": [ + { + "fallback": "Plain-text summary of the attachment.", + "color": "<% if (success) { %>good<% } else { %>danger<%} %>", + "author_name": "nf-core/rnadeseq ${version} - ${runName}", + "author_icon": "https://www.nextflow.io/docs/latest/_static/favicon.ico", + "text": "<% if (success) { %>Pipeline completed successfully!<% } else { %>Pipeline completed with errors<% } %>", + "fields": [ + { + "title": "Command used to launch the workflow", + "value": "```${commandLine}```", + "short": false + } + <% + if (!success) { %> + , + { + "title": "Full error message", + "value": "```${errorReport}```", + "short": false + }, + { + "title": "Pipeline configuration", + "value": "<% out << summary.collect{ k,v -> k == "hook_url" ? "_${k}_: (_hidden_)" : ( ( v.class.toString().contains('Path') || ( v.class.toString().contains('String') && v.contains('/') ) ) ? "_${k}_: `${v}`" : (v.class.toString().contains('DateTime') ? ("_${k}_: " + v.format(java.time.format.DateTimeFormatter.ofLocalizedDateTime(java.time.format.FormatStyle.MEDIUM))) : "_${k}_: ${v}") ) }.join(",\n") %>", + "short": false + } + <% } + %> + ], + "footer": "Completed at <% out << dateComplete.format(java.time.format.DateTimeFormatter.ofLocalizedDateTime(java.time.format.FormatStyle.MEDIUM)) %> (duration: ${duration})" + } + ] +} diff --git a/conf/base.config b/conf/base.config new file mode 100644 index 00000000..b709a294 --- /dev/null +++ b/conf/base.config @@ -0,0 +1,62 @@ +/* +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ + nf-core/rnadeseq Nextflow base config file +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ + A 'blank slate' config file, appropriate for general use on most high performance + compute environments. Assumes that all software is installed and available on + the PATH. Runs in `local` mode - all jobs will be run on the logged in environment. +---------------------------------------------------------------------------------------- +*/ + +process { + + // TODO nf-core: Check the defaults for all processes + cpus = { check_max( 1 * task.attempt, 'cpus' ) } + memory = { check_max( 6.GB * task.attempt, 'memory' ) } + time = { check_max( 4.h * task.attempt, 'time' ) } + + errorStrategy = { task.exitStatus in ((130..145) + 104) ? 'retry' : 'finish' } + maxRetries = 1 + maxErrors = '-1' + + // Process-specific resource requirements + // NOTE - Please try and re-use the labels below as much as possible. + // These labels are used and recognised by default in DSL2 files hosted on nf-core/modules. + // If possible, it would be nice to keep the same label naming convention when + // adding in your local modules too. + // TODO nf-core: Customise requirements for specific processes. + // See https://www.nextflow.io/docs/latest/config.html#config-process-selectors + withLabel:process_single { + cpus = { check_max( 1 , 'cpus' ) } + memory = { check_max( 6.GB * task.attempt, 'memory' ) } + time = { check_max( 4.h * task.attempt, 'time' ) } + } + withLabel:process_low { + cpus = { check_max( 2 * task.attempt, 'cpus' ) } + memory = { check_max( 12.GB * task.attempt, 'memory' ) } + time = { check_max( 4.h * task.attempt, 'time' ) } + } + withLabel:process_medium { + cpus = { check_max( 6 * task.attempt, 'cpus' ) } + memory = { check_max( 36.GB * task.attempt, 'memory' ) } + time = { check_max( 8.h * task.attempt, 'time' ) } + } + withLabel:process_high { + cpus = { check_max( 12 * task.attempt, 'cpus' ) } + memory = { check_max( 72.GB * task.attempt, 'memory' ) } + time = { check_max( 16.h * task.attempt, 'time' ) } + } + withLabel:process_long { + time = { check_max( 20.h * task.attempt, 'time' ) } + } + withLabel:process_high_memory { + memory = { check_max( 200.GB * task.attempt, 'memory' ) } + } + withLabel:error_ignore { + errorStrategy = 'ignore' + } + withLabel:error_retry { + errorStrategy = 'retry' + maxRetries = 2 + } +} diff --git a/conf/igenomes.config b/conf/igenomes.config new file mode 100644 index 00000000..3f114377 --- /dev/null +++ b/conf/igenomes.config @@ -0,0 +1,440 @@ +/* +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ + Nextflow config file for iGenomes paths +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ + Defines reference genomes using iGenome paths. + Can be used by any config that customises the base path using: + $params.igenomes_base / --igenomes_base +---------------------------------------------------------------------------------------- +*/ + +params { + // illumina iGenomes reference file paths + genomes { + 'GRCh37' { + fasta = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Annotation/README.txt" + mito_name = "MT" + macs_gsize = "2.7e9" + blacklist = "${projectDir}/assets/blacklists/GRCh37-blacklist.bed" + } + 'GRCh38' { + fasta = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Annotation/Genes/genes.bed" + mito_name = "chrM" + macs_gsize = "2.7e9" + blacklist = "${projectDir}/assets/blacklists/hg38-blacklist.bed" + } + 'CHM13' { + fasta = "${params.igenomes_base}/Homo_sapiens/UCSC/CHM13/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Homo_sapiens/UCSC/CHM13/Sequence/BWAIndex/" + bwamem2 = "${params.igenomes_base}/Homo_sapiens/UCSC/CHM13/Sequence/BWAmem2Index/" + gtf = "${params.igenomes_base}/Homo_sapiens/NCBI/CHM13/Annotation/Genes/genes.gtf" + gff = "ftp://ftp.ncbi.nlm.nih.gov/genomes/all/GCF/009/914/755/GCF_009914755.1_T2T-CHM13v2.0/GCF_009914755.1_T2T-CHM13v2.0_genomic.gff.gz" + mito_name = "chrM" + } + 'GRCm38' { + fasta = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Annotation/README.txt" + mito_name = "MT" + macs_gsize = "1.87e9" + blacklist = "${projectDir}/assets/blacklists/GRCm38-blacklist.bed" + } + 'TAIR10' { + fasta = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Annotation/README.txt" + mito_name = "Mt" + } + 'EB2' { + fasta = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Annotation/README.txt" + } + 'UMD3.1' { + fasta = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Annotation/README.txt" + mito_name = "MT" + } + 'WBcel235' { + fasta = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Annotation/Genes/genes.bed" + mito_name = "MtDNA" + macs_gsize = "9e7" + } + 'CanFam3.1' { + fasta = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Annotation/README.txt" + mito_name = "MT" + } + 'GRCz10' { + fasta = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Annotation/Genes/genes.bed" + mito_name = "MT" + } + 'BDGP6' { + fasta = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Annotation/Genes/genes.bed" + mito_name = "M" + macs_gsize = "1.2e8" + } + 'EquCab2' { + fasta = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Annotation/README.txt" + mito_name = "MT" + } + 'EB1' { + fasta = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Annotation/README.txt" + } + 'Galgal4' { + fasta = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Annotation/Genes/genes.bed" + mito_name = "MT" + } + 'Gm01' { + fasta = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Annotation/README.txt" + } + 'Mmul_1' { + fasta = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Annotation/README.txt" + mito_name = "MT" + } + 'IRGSP-1.0' { + fasta = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Annotation/Genes/genes.bed" + mito_name = "Mt" + } + 'CHIMP2.1.4' { + fasta = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Annotation/README.txt" + mito_name = "MT" + } + 'Rnor_5.0' { + fasta = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_5.0/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_5.0/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_5.0/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_5.0/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_5.0/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_5.0/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_5.0/Annotation/Genes/genes.bed" + mito_name = "MT" + } + 'Rnor_6.0' { + fasta = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Annotation/Genes/genes.bed" + mito_name = "MT" + } + 'R64-1-1' { + fasta = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Annotation/Genes/genes.bed" + mito_name = "MT" + macs_gsize = "1.2e7" + } + 'EF2' { + fasta = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Annotation/README.txt" + mito_name = "MT" + macs_gsize = "1.21e7" + } + 'Sbi1' { + fasta = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Annotation/README.txt" + } + 'Sscrofa10.2' { + fasta = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Annotation/README.txt" + mito_name = "MT" + } + 'AGPv3' { + fasta = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Annotation/Genes/genes.bed" + mito_name = "Mt" + } + 'hg38' { + fasta = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Annotation/Genes/genes.bed" + mito_name = "chrM" + macs_gsize = "2.7e9" + blacklist = "${projectDir}/assets/blacklists/hg38-blacklist.bed" + } + 'hg19' { + fasta = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Annotation/README.txt" + mito_name = "chrM" + macs_gsize = "2.7e9" + blacklist = "${projectDir}/assets/blacklists/hg19-blacklist.bed" + } + 'mm10' { + fasta = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Annotation/README.txt" + mito_name = "chrM" + macs_gsize = "1.87e9" + blacklist = "${projectDir}/assets/blacklists/mm10-blacklist.bed" + } + 'bosTau8' { + fasta = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Annotation/Genes/genes.bed" + mito_name = "chrM" + } + 'ce10' { + fasta = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Annotation/README.txt" + mito_name = "chrM" + macs_gsize = "9e7" + } + 'canFam3' { + fasta = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Annotation/README.txt" + mito_name = "chrM" + } + 'danRer10' { + fasta = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Annotation/Genes/genes.bed" + mito_name = "chrM" + macs_gsize = "1.37e9" + } + 'dm6' { + fasta = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Annotation/Genes/genes.bed" + mito_name = "chrM" + macs_gsize = "1.2e8" + } + 'equCab2' { + fasta = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Annotation/README.txt" + mito_name = "chrM" + } + 'galGal4' { + fasta = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Annotation/README.txt" + mito_name = "chrM" + } + 'panTro4' { + fasta = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Annotation/README.txt" + mito_name = "chrM" + } + 'rn6' { + fasta = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Annotation/Genes/genes.bed" + mito_name = "chrM" + } + 'sacCer3' { + fasta = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/BismarkIndex/" + readme = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Annotation/README.txt" + mito_name = "chrM" + macs_gsize = "1.2e7" + } + 'susScr3' { + fasta = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/BWAIndex/version0.6.0/" + bowtie2 = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Annotation/README.txt" + mito_name = "chrM" + } + } +} diff --git a/conf/modules.config b/conf/modules.config new file mode 100644 index 00000000..d203d2b6 --- /dev/null +++ b/conf/modules.config @@ -0,0 +1,34 @@ +/* +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ + Config file for defining DSL2 per module options and publishing paths +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ + Available keys to override module options: + ext.args = Additional arguments appended to command in module. + ext.args2 = Second set of arguments appended to command in module (multi-tool modules). + ext.args3 = Third set of arguments appended to command in module (multi-tool modules). + ext.prefix = File name prefix for output files. +---------------------------------------------------------------------------------------- +*/ + +process { + + publishDir = [ + path: { "${params.outdir}/${task.process.tokenize(':')[-1].tokenize('_')[0].toLowerCase()}" }, + mode: params.publish_dir_mode, + saveAs: { filename -> filename.equals('versions.yml') ? null : filename } + ] + + withName: FASTQC { + ext.args = '--quiet' + } + + withName: 'MULTIQC' { + ext.args = { params.multiqc_title ? "--title \"$params.multiqc_title\"" : '' } + publishDir = [ + path: { "${params.outdir}/multiqc" }, + mode: params.publish_dir_mode, + saveAs: { filename -> filename.equals('versions.yml') ? null : filename } + ] + } + +} diff --git a/conf/test.config b/conf/test.config new file mode 100644 index 00000000..8662d422 --- /dev/null +++ b/conf/test.config @@ -0,0 +1,29 @@ +/* +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ + Nextflow config file for running minimal tests +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ + Defines input files and everything required to run a fast and simple pipeline test. + + Use as follows: + nextflow run nf-core/rnadeseq -profile test, --outdir + +---------------------------------------------------------------------------------------- +*/ + +params { + config_profile_name = 'Test profile' + config_profile_description = 'Minimal test dataset to check pipeline function' + + // Limit resources so that this can run on GitHub Actions + max_cpus = 2 + max_memory = '6.GB' + max_time = '6.h' + + // Input data + // TODO nf-core: Specify the paths to your test data on nf-core/test-datasets + // TODO nf-core: Give any required params for the test so that command line flags are not needed + input = params.pipelines_testdata_base_path + 'viralrecon/samplesheet/samplesheet_test_illumina_amplicon.csv' + + // Genome references + genome = 'R64-1-1' +} diff --git a/conf/test_full.config b/conf/test_full.config new file mode 100644 index 00000000..f6e6cec8 --- /dev/null +++ b/conf/test_full.config @@ -0,0 +1,24 @@ +/* +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ + Nextflow config file for running full-size tests +~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ + Defines input files and everything required to run a full size pipeline test. + + Use as follows: + nextflow run nf-core/rnadeseq -profile test_full, --outdir + +---------------------------------------------------------------------------------------- +*/ + +params { + config_profile_name = 'Full test profile' + config_profile_description = 'Full test dataset to check pipeline function' + + // Input data for full size test + // TODO nf-core: Specify the paths to your full test data ( on nf-core/test-datasets or directly in repositories, e.g. SRA) + // TODO nf-core: Give any required params for the test so that command line flags are not needed + input = params.pipelines_testdata_base_path + 'viralrecon/samplesheet/samplesheet_full_illumina_amplicon.csv' + + // Genome references + genome = 'R64-1-1' +} diff --git a/docs/README.md b/docs/README.md new file mode 100644 index 00000000..dc052cc1 --- /dev/null +++ b/docs/README.md @@ -0,0 +1,10 @@ +# nf-core/rnadeseq: Documentation + +The nf-core/rnadeseq documentation is split into the following pages: + +- [Usage](usage.md) + - An overview of how the pipeline works, how to run it and a description of all of the different command-line flags. +- [Output](output.md) + - An overview of the different results produced by the pipeline and how to interpret them. + +You can find a lot more documentation about installing, configuring and running nf-core pipelines on the website: [https://nf-co.re](https://nf-co.re) diff --git a/docs/images/mqc_fastqc_adapter.png b/docs/images/mqc_fastqc_adapter.png new file mode 100755 index 0000000000000000000000000000000000000000..361d0e47acfb424dea1f326590d1eb2f6dfa26b5 GIT binary patch literal 23458 zcmeFZ2UJtryD!S#x<#o93es(Ww4k)maRbte0-+a?-g^xY-3myTE`8G_KvA54)F1tn})nJ5u%TA4Y;^!^{48eL_}p#q-Umo0M|F1 z74+PQh^X8N|9_jcWbq~ zzn+tZC9B75nKdz=gQ8wo9GJ$P{D~3knlI_`-PRhCw34f1oYDLr^;oEbgxa#A^J%*2 z>FfDE*(~JzKFs$t_oeLz))qDU?s}%Q?7b~3Y;lUi^Oy-2@3g?joA4Wkgb6-2=ih*jub)~7yZ`T=L=Z`B`{1jhkB-iSjea94&Eo9A zxN59pv1p_}RO1>EC^q}Z2)ZI;b7JV_x4lMr=Bker2+EK;8~!;JO7re*@ZkDmoV878S*N^yX(F@U1yqt?Is3nnV>7}#(5pk`V3C) 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Output + +## Introduction + +This document describes the output produced by the pipeline. Most of the plots are taken from the MultiQC report, which summarises results at the end of the pipeline. + +The directories listed below will be created in the results directory after the pipeline has finished. All paths are relative to the top-level results directory. + + + +## Pipeline overview + +The pipeline is built using [Nextflow](https://www.nextflow.io/) and processes data using the following steps: + +- [FastQC](#fastqc) - Raw read QC +- [MultiQC](#multiqc) - Aggregate report describing results and QC from the whole pipeline +- [Pipeline information](#pipeline-information) - Report metrics generated during the workflow execution + +### FastQC + +