Header Leaderboard Ad

Collapse

samtools mpileup did not call variant

Collapse

Announcement

Collapse

SEQanswers June Challenge Has Begun!

The competition has begun! We're giving away a $50 Amazon gift card to the member who answers the most questions on our site during the month. We want to encourage our community members to share their knowledge and help each other out by answering questions related to sequencing technologies, genomics, and bioinformatics. The competition is open to all members of the site, and the winner will be announced at the beginning of July. Best of luck!

For a list of the official rules, visit (https://www.seqanswers.com/forum/sit...wledge-and-win)
See more
See less
X
 
  • Filter
  • Time
  • Show
Clear All
new posts

  • samtools mpileup did not call variant

    I used samtools mpileup and bcftools to call variants in my RNA-Seq data. At one locus, I have 10 reads that are different from the ref sequence (DP4=32,21,5,5), but bcftools did not call this variant.

    3 13812321 . T . 47 . DP=65;VDB=0.0190;AF1=0;AC1=0;DP4=32,21,5,5;MQ=20;FQ=-44;PV4=0.73,1,1,1 PL: DP:SP 0:63:1

    I tried to change the -p value from the default 0.5 to 0.6, 0.7, 0.8 and 0.9 but still none of them called this variant. I understand that 10 reads in 65 reads is a low fraction, but is there anyway to make bcftools less stringent? I know I can just print out info for every locus and do my own filtering but I was just wondering if there are other parameters of bcftool tat I can modify. Thanks!

  • #2
    Isn't Mapq=20 rather too low to confidently take the alignment at that position?

    Comment


    • #3
      Thank you for the reply. No, I dont think MQ=20 was the problem. Some of the called variants in my run had MQ as low as 16. Any other thoughts?

      Comment

      Latest Articles

      Collapse

      ad_right_rmr

      Collapse

      News

      Collapse

      Topics Statistics Last Post
      Started by seqadmin, Yesterday, 10:20 AM
      0 responses
      9 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 06-07-2023, 07:14 AM
      0 responses
      13 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 06-06-2023, 01:08 PM
      0 responses
      13 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 06-01-2023, 08:56 PM
      0 responses
      166 views
      0 likes
      Last Post seqadmin  
      Working...
      X