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  • liying
    Member
    • Sep 2010
    • 11

    #1

    How samtools computes AF1?

    Hi,Sorry to ask basic.

    I get such vcf by smtools mpileup, But can't understand it.

    scaffold_1 8463589 . T C 47 . DP=5;VDB=0.0520;AF1=0.500;AFE=0.500;DP4=0,0,3,2;MQ=38 PL:GT:GQ 92,15,92:0/1:77

    The ref is T, all my 5 reads are C at this site. So how the AF1 (site allele frequency of the strongest non-reference allele) is 0.5 ?
    How samtools computes AF1?

    PS: I didn't find any explain about AFE. what is it?

    Thanks!
  • liying
    Member
    • Sep 2010
    • 11

    #2
    It turns out be a version error. We didn't change $PATH of bcftools when updata samtools.
    Now the result is

    scaffold_1 8463589 . T C 59.3 . DP=5;VDB=0.0520;AF1=1;AC1=2;DP4=0,0,3,2;MQ=38;FQ=-42 GT:PL:GQ 1/1:92,15,0:27

    AF1 look like alright. But what are VDB and AC1? I didn't find them in samtools, mpileup and VCF document.
    I'm using samtools 0.1.18. Are them some new in this version?

    Comment

    • Heisman
      Senior Member
      • Dec 2010
      • 534

      #3
      Both of those are described (briefly) in the header section of the SNP files that are generated.

      ##INFO=<ID=AC1,Number=1,Type=Float,Description="Max-likelihood estimate of the first ALT allele count (no HWE assumption)">

      ##INFO=<ID=VDB,Number=1,Type=Float,Description="Variant Distance Bias">

      Comment

      • liying
        Member
        • Sep 2010
        • 11

        #4
        Heisman,thank you very much!
        My vcf file don't have any header. Could you post you command?

        My command:
        samtools mpileup -A -q 20 -Q 13 -C 50 -uf Egrandis_162.fa 2.rmdup.bam | bcftools view -bvcg - >2.RawVar.bcf 2>mpileup.log
        bcftools view 2.RawVar.bcf | vcfutils.pl varFilter -Q 30 -d 2 - > 2.d2Q30.FilterVar.vcf

        Comment

        • Heisman
          Senior Member
          • Dec 2010
          • 534

          #5
          samtools-0.1.18/samtools mpileup -AB -ugf [Reference_Sequence] [aligned_file] | samtools-0.1.18/bcftools/bcftools view -bvcg -> [Intermediate_File] &

          samtools-0.1.18/bcftools/bcftools view [Intermediate_File] | samtools-0.1.18/bcftools/vcfutils.pl varFilter -D99999 > [Variant_Output] &

          Comment

          • liying
            Member
            • Sep 2010
            • 11

            #6
            Heisman,thanks!

            why use -g and -u together? We used piping, so it should be -u.

            samtools mpileup
            -g Compute genotype likelihoods and output them in the binary call format (BCF).
            -u Similar to -g except that the output is uncompressed BCF, which is preferred for piping.

            Well,I can't see why my file missing header.
            Last edited by liying; 09-26-2011, 09:13 PM.

            Comment

            • jy123
              Junior Member
              • Nov 2010
              • 8

              #7
              Hi,

              I have a problem to understand the following result:
              chr4 114612702 . G A 52 . DP=29;VDB=0.0218;AF1=1;AC1=8;DP4=6,4,10,9

              How come that AF1=1 with ref and alt reads number having no big difference?

              Comment

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