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ksilnoaa authored Aug 10, 2023
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44 changes: 24 additions & 20 deletions README.md
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Expand Up @@ -68,20 +68,24 @@ Before you download the Tourmaline commands and directory structure from GitHub,

#### Option 1: Native installation

To run Tourmaline natively on a Mac (Intel) or Linux system, start with a Conda installation of QIIME 2 (for Linux, change "osx" to "linux"):
To run Tourmaline natively on a Mac (Intel) or Linux system, start with a Conda installation of Snakemake v7.30.1. We recommend using [Miniconda with a python >= 3.8](https://docs.conda.io/en/latest/miniconda.html):

```bash
wget https://data.qiime2.org/distro/core/qiime2-2023.2-py38-osx-conda.yml
# make sure you are using the most updated conda
conda update conda
conda env create -n qiime2-2023.2 --file qiime2-2023.2-py38-osx-conda.yml
conda install -c conda-forge -c bioconda snakemake=7.30.1
```

Then install QIIME 2 with conda (**for Linux, change "osx" to "linux"**):

```bash
wget https://data.qiime2.org/distro/core/qiime2-2023.5-py38-osx-conda.yml
conda env create -n qiime2-2023.5 --file qiime2-2023.5-py38-osx-conda.yml
```

Activate the environment and install the other Conda- or PIP-installable dependencies:

```
conda activate qiime2-2023.2
conda install -c conda-forge -c bioconda snakemake biopython muscle clustalo tabulate
conda activate qiime2-2023.5
conda install -c conda-forge -c bioconda biopython muscle clustalo tabulate
conda install -c conda-forge deicode
pip install empress
qiime dev refresh-cache
Expand All @@ -95,9 +99,9 @@ Follow these instructions for Macs with M1/M2 chips.
**First, set your Terminal application to run in [Rosetta mode](https://academy.bigbinary.com/learn-rubyonrails/setting-up-macos).**

```bash
wget https://data.qiime2.org/distro/core/qiime2-2023.2-py38-osx-conda.yml
CONDA_SUBDIR=osx-64 conda env create -n qiime2-2023.2 --file qiime2-2023.2-py38-osx-conda.yml
conda activate qiime2-2023.2
wget https://data.qiime2.org/distro/core/qiime2-2023.5-py38-osx-conda.yml
CONDA_SUBDIR=osx-64 conda env create -n qiime2-2023.5 --file qiime2-2023.5-py38-osx-conda.yml
conda activate qiime2-2023.5
conda config --env --set subdir osx-64
```

Expand Down Expand Up @@ -154,8 +158,8 @@ Download reference database sequence and taxonomy files, named `refseqs.qza` and

```bash
cd tourmaline/01-imported
wget https://data.qiime2.org/2023.2/common/silva-138-99-seqs-515-806.qza
wget https://data.qiime2.org/2023.2/common/silva-138-99-tax-515-806.qza
wget https://data.qiime2.org/2023.5/common/silva-138-99-seqs-515-806.qza
wget https://data.qiime2.org/2023.5/common/silva-138-99-tax-515-806.qza
ln -s silva-138-99-seqs-515-806.qza refseqs.qza
ln -s silva-138-99-tax-515-806.qza reftax.qza
```
Expand Down Expand Up @@ -193,12 +197,12 @@ Now edit, replace, or store the required input files as described here:

Shown here is the DADA2 paired-end workflow. See the Wiki's [Run](https://github.com/aomlomics/tourmaline/wiki/4-Run) page for complete instructions on all steps, denoising methods, and filtering modes.

Note that any of the commands below can be run with various options, including `--printshellcmds` to see the shell commands being executed and `--dryrun` to display which rules would be run but not execute them. To generate a graph of the rules that will be run from any Snakemake command, see the section "Directed acyclic graph (DAG)" on the [Run](https://github.com/aomlomics/tourmaline/wiki/4-Run) page.
Note that any of the commands below can be run with various options, including `--printshellcmds` to see the shell commands being executed and `--dryrun` to display which rules would be run but not execute them. To generate a graph of the rules that will be run from any Snakemake command, see the section "Directed acyclic graph (DAG)" on the [Run](https://github.com/aomlomics/tourmaline/wiki/4-Run) page. **Always include the --use-conda option.**

From the `tourmaline` directory (which you may rename), run Snakemake with the *denoise* rule as the target, changing the number of cores to match your machine:

```bash
snakemake dada2_pe_denoise --cores 4
snakemake --use-conda dada2_pe_denoise --cores 4
```

Pausing after the *denoise* step allows you to make changes before proceeding:
Expand All @@ -212,19 +216,19 @@ Pausing after the *denoise* step allows you to make changes before proceeding:
Continue the workflow without filtering (for now). If you are satisfied with your parameters and files, run the *taxonomy* rule (for unfiltered data):

```bash
snakemake dada2_pe_taxonomy_unfiltered --cores 4
snakemake --use-conda dada2_pe_taxonomy_unfiltered --cores 4
```

Next, run the *diversity* rule (for unfiltered data):

```bash
snakemake dada2_pe_diversity_unfiltered --cores 4
snakemake --use-conda dada2_pe_diversity_unfiltered --cores 4
```

Finally, run the *report* rule (for unfiltered data):

```bash
snakemake dada2_pe_report_unfiltered --cores 4
snakemake --use-conda dada2_pe_report_unfiltered --cores 4
```

#### Filtered mode
Expand All @@ -239,19 +243,19 @@ After viewing the *unfiltered* results—the taxonomy summary and taxa barplot,
Now we are ready to filter the representative sequences and feature table, generate new summaries, and generate a new taxonomy bar plot, by running the *taxonomy* rule (for filtered data):

```bash
snakemake dada2_pe_taxonomy_filtered --cores 4
snakemake --use-conda dada2_pe_taxonomy_filtered --cores 4
```

Next, run the *diversity* rule (for filtered data):

```bash
snakemake dada2_pe_diversity_filtered --cores 4
snakemake --use-conda dada2_pe_diversity_filtered --cores 4
```

Finally, run the *report* rule (for filtered data):

```bash
snakemake dada2_pe_report_filtered --cores 1
snakemake --use-conda dada2_pe_report_filtered --cores 1
```

### View output
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