Missing gentoypes despite high number of reads

Dear GATK-team

GenotypeGVCFs, sets all genotypes as missing for a large part of a gene I am interested in, even though there seem to be lots of reads in all individuals for those sites. I have the same problem if I am using versions 3.5 or 3.7. It also makes no difference if HaplotypeCaller was run with -ERC BP_Resolution or the standard GVCF, or if I run GenotypeGVCFs on just one individual or many at once. Could it be related to the fact that this is a mitochondrial region and thus has higher sequencing depth and theoretically no heterozygote positions?

Here is an example of the vcf file produced by GenotypeGVCFs:
chrM 16758 . A . 67.16 . DP=14015 GT:AD:DP ./.:444,9:453 ./.:257,0:257 (...)

In the g.vcf file of the first individual this site looks like this (the 9 alternative allele reads are likely contamination):
chrM 16758 . A . . . GT:AD:DP:GQ:PL 0/0:444,9:453:0:0,0,0

And in the second individual g.vcf:
chrM 16758 . A . . . GT:AD:DP:GQ:PL 0/0:257,0:257:0:0,0,0

Any advice would be highly appreciated.

Many thanks!

Answers

  • SheilaSheila Broad InstituteMember, Broadie, Moderator

    @JoanaM
    Hi,

    The reason they are being set to no-calls is that the GQs are all 0. Notice the PLs are all 0. Can you post IGV screenshots of the original BAM file and bamout file? Please include ~300 bases before and after the site of interest.

    Thanks,
    Sheila

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