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Depth Reporting in DP and AD changes when VariantAnnotator run

Hello,

I am trying to filter some of my high-coverage samples based on a minimum depth and have found that the value stored in the DP INFO field and the AD genotype tag changes depending on whether or not I have run VariantAnnotator. The call I have used for VariantAnnotator is:

java -jar GenomeAnalysisTK.jar -T VariantAnnotator -R ucsc.hg19.fasta -I example.bam --variant example.raw.vcf --out example.annotated.vcf -G StandardAnnotation -L example.raw.vcf -rf BadCigar -dcov 15000

Here are the differences for some test cases with HaplotypeCaller:

No MarkDuplicates, did IndelRealigner & BQSR, nightly build 12/04/2013

Annotated: DP=2745, AD=4,2729

Raw: DP=957, AD=1,907

MarkDuplicates, IndelRealigner and BQSR, nightly build 12/04/2013

Annotated: DP=20, AD=0,20

Raw: DP=10, AD=0,8

Raw BAM, nightly build 12/04/2013

Annotated: DP=2745,AD=4,2729

Raw: DP=868, AD=1,864

Raw BAM, version 2.4-9

Annotated: DP=2745, AD=4,2729

Raw: DP=616, AD=1,611

I suspect what is happening here is that VariantAnnotator is taking the depth information from the provided BAM and replacing the depth information reported by the variant caller. Anyway, just wondering- which value is a better reflection of the depth used to make a given variant call? (i.e. which could I use in hard filtering?)

Thanks for your help!

Best Answer

Answers

  • nhvanlienhvanlie Member

    Great, thank you for confirming! In this case, I suspect we are seeing more genuine duplication (because of small, targeted areas and high coverage) than PCR duplication, so I'll have to do more testing to determine whether I should run MarkDuplicates.

  • Geraldine_VdAuweraGeraldine_VdAuwera admin Cambridge, MAMember, Administrator, Broadie admin

    That's a fair point, if you know the duplicates are legit. From those values it looks like the results are consistent either way.

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