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featurecount command to extract read count from bam files

I m using this featurecount command to extract read count from my bam files , is it the correct one im using or there is some issue with my command .

featureCounts -T 40 -p -A  -s -f -O -t exon -g gene_id -a /home/punit/ERCCgencode.v21.annotation.gtf -o /home/punit/FCOUNT/newhl60.txt ~/bamfiles/WT1.bam ~/bamfiles/WT2.bam ~/bamfiles/AT1.bam ~/bamfiles/AT2.bam ~/bamfiles/VD1.bam ~/bamfiles/VD2.bam
rna-seq
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updated by Devon Ryan Devon Ryan • written by 1769mkc 1769mkc

1 answer

  • -A takes a file with chromosome name aliases, which you're not providing and likely don't need.
  • -s takes a number indicating what sort of strand specificity you want. You likely want -s 2.
  • If you want to use these in edgeR/DESeq2/etc., don't use -O.
    • You probably don't want -f.

In general, run commands first and spot-check the results to see if anything went wrong. Also, start using new commands/tools by using the defaults.

• 0 views • link
written by Devon Ryan Devon Ryan

yes Im using the output for deseq2

• 0 views • link
written by 1769mkc 1769mkc

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