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NGSBioAfrica

Module 1: Unix

Reading and Editing files SED AWK BASH Scripting

Module 2: NGS Technologies

1st generation-sangersequencing(short reads) 2nd generation-illumina,ion torrent/thermo fischer(short reads) 3rd generation-nanopore,pacbio SMRT (long reads)

Long and short reads can complement each other

  • Long reads improve genome assembly
  • Assembly can be corrected by short reads
  • Long reads bridge regions with high error rate
  • Short reads can correct base calls of long reads

High Throughput Sequencing offeres a broad range of applications

  • Discover variants
  • Population studies
  • Analyze clinical isolates
  • Study rare diseases
  • Characterize the genome
  • Characterize DNA-Protein(TF)interactions

Challenges of HT Sequencing

  • Data collection-extensive uses of resources,sampling accuracy
  • Data analysis and interpretation
  • Analytical/Computational challenges

Module 3: NGS Data Analysis Overview and Data Formats

Obtaining data to be used

cd course_data/data_formats/data unzip SRR957824 cd SRR957824

Quality check

fastqc SRR957824_1.fastq.gz fastqc SRR957824_2.fastq.gz or fastqc SRR957824_1.fastq.gz SRR957824_2.fastq.gz or fastqc *.fastq.gz (using a wildcard)

multiqc ./

#to view the quality reports firefox SRR957824_1.fastq.html firefox SRR957824_2.fastq.html firefox multiqc*.html