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Variant Discovery in High-Throughput Sequencing Data

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Developed in the Data Sciences Platform at the Broad Institute, the toolkit offers a wide variety of tools with a primary focus on variant discovery and genotyping. Its powerful processing engine and high-performance computing features make it capable of taking on projects of any size. Learn more

--minimum-allele-fraction does not work, vcf output is not filtered properly

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    David Benjamin

    Hiral Desai Mutect2 does not do filtering; that is the job of FilterMutectCalls.  Also, this argument is not an AF threshold for filtering variants, see the command line help docs for details.  In any case, we do not recommend hard filtering in this manner.

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    Hiral Desai

    thank you, why is the argument available then? I also tried it for FilterMutectCalls and it does not work as intended. The expected results don't filter according to my set AF or AD.

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    David Benjamin

    We do not recommend hard filtering by AF and AD.  We recommend using the defaults for Mutect2 and FilterMutectCalls except in exceptional circumstances.

    The argument works as intended but the intent is not filtering.  It is a very technical detail of the mitochondrial GVCF mode that we needed for the gnomAD mitochondria project.

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    Hiral Desai

    I'm using it for viral variant calling and in my circumstance, I need to set more criteria based on ground truth data we've been studying. The command line options are available whether it's FilterMutectCalls or VariantFiltration but even if I add a filter flag such as AD  < 3 and provide the expression PASS OR lowReads, the VCF output does not format accordingly.

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    David Benjamin

    Viral variant calling could, possibly, be an exceptional circumstance.  I'm not sure what argument you are referring to in FilterMutectCalls, but I believe that the VariantFiltration command would be something like:

    gatk VariantFiltration \
    -R reference.fasta \
    -V input.vcf \
    -O output.vcf \
    --filter-name "lowReads" \
    --filter-expression "AD < 3"
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