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  • Hard trims FASTQ, if optionally specified (fastx_trimmer)
  • Performs the global or local alignment (here, a PE alignment)
    • BWA globalbwa aln the R1 and R2 separately, then bwa sampe to produce a SAM file
    • BWA local: call bwa mem with both R1 and R2 to produce a SAM file
    • Bowtie2 global: call bowtie2 in its default global (end--global with to-end) mode on both R1 and R2 to produce a SAM file
    • Bowtie2 local: call bowtie2 --local with both R1 and R2 to produce a SAM file
  • Converts SAM to BAM (samtools view)
  • Sorts the BAM (samtools sort)
  • Marks duplicates (Picard MarkDuplicates)
  • Indexes the sorted, duplicate-marked BAM (samtools index)
  • Gathers statistics (samtools idxstats, samtools flagstat, plus a custom statistics script of Anna's)
  • Removes intermediate files

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