Haploid genomes

gilgigilgi Posts: 19Member
edited July 2012 in Ask the GATK team

Dear GATK team,

I know that in the past GATK was not suitable for haploid genomes.
I wanted to ask if this possibly changed since then - and whether it is possible to use GATK for haploid genomes.

Thanks a lot,
Gilgi

Post edited by Carneiro on

Best Answer

  • Mark_DePristoMark_DePristo Posts: 153Administrator, GATK Dev admin
    Answer ✓

    The ug in gatk2 can call haploid sequence natively now. You just set ploidy to 1.

    --
    Mark A. DePristo, Ph.D.
    Co-Director, Medical and Population Genetics
    Broad Institute of MIT and Harvard

Answers

  • Mark_DePristoMark_DePristo Posts: 153Administrator, GATK Dev admin
    Answer ✓

    The ug in gatk2 can call haploid sequence natively now. You just set ploidy to 1.

    --
    Mark A. DePristo, Ph.D.
    Co-Director, Medical and Population Genetics
    Broad Institute of MIT and Harvard

  • gilgigilgi Posts: 19Member

    Thanks! This is great news! I'll try to work with it.

  • gilgigilgi Posts: 19Member

    Hi,
    We tried to download gatk2 from here:
    http://www.broadinstitute.org/gatk/download

    But after download saw that the version doesn't seem to be 2 but: version 1.6-596-g3b9929c

    Is this the correct version?

    Thanks,
    Gilgi

  • ebanksebanks Broad InstitutePosts: 687Member, Administrator, GATK Dev, Broadie, Moderator, DSDE Dev, GP Member admin

    The 2.0 release should be coming out later today.

    Eric Banks, PhD -- Senior Group Leader, MPG Analysis, Broad Institute of Harvard and MIT

  • gilgigilgi Posts: 19Member

    Thanks a lot. I downloaded the gatk 2.0.

    When trying to use the UnifiedGenotyper with --sample_ploidy 1 I get an error:
    MESSAGE: Incorrect genotype calculation model chosen. Only [POOLSNP|POOLINDEL|POOLBOTH] supported with this walker if sample ploidy != 2

    What does this mean? My data isn't pool, I have individual (barcoded) haploid sequenced strains.
    I tried to add:
    --genotype_likelihoods_model POOLBOTH

    But then I get:
    MESSAGE: Incorrect AF Calculation model. Only POOL model supported if sample ploidy != 2

    I tried to look for the answer in the guide - without success.
    Can you help please?

  • gilgigilgi Posts: 19Member

    Thanks a lot!!!

  • hillihilli Posts: 1Member

    Ive used the -pnrm POOL option but still getting the same error as gilgi. So I have

    java -Xmx30g -jar /usr/local/gatk2/GenomeAnalysisTK.jar -T UnifiedGenotyper -R ref -I bam -I bam -pnrm POOL -polidy 1 -o vcf

    Any help?
    Ali

  • ebanksebanks Broad InstitutePosts: 687Member, Administrator, GATK Dev, Broadie, Moderator, DSDE Dev, GP Member admin

    We have changed the arguments so that they more accurately reflect what they are doing:
    http://www.broadinstitute.org/gatk/gatkdocs/org_broadinstitute_sting_gatk_walkers_genotyper_UnifiedGenotyper.html#--genotype_likelihoods_model
    So you'll want e.g. -pnrm GeneralPloidySNP

    Eric Banks, PhD -- Senior Group Leader, MPG Analysis, Broad Institute of Harvard and MIT

  • flowflow Posts: 14Member

    With v2.0-39 I had been using -ploidy 1 -pnrm POOL -glm POOLSNP. Can you confirm that for calling of snps/indels in haploid genomes as of v2.1 would now be -ploidy 1 -pnrm EXACT -glm GeneralPloidySNP. Is the -pnrm POOL option now defunct?

  • delangeldelangel Posts: 71GATK Dev mod

    You just need -ploidy 1. "-pnrm EXACT" will work but there's no other option :). "-glm GeneralPloidySNP" will not work - you need either SNP, INDEL or BOTH.

  • flowflow Posts: 14Member

    Thanks for your reply. Under what circumstance should -glm GeneralPloidySNP/GeneralPloidyINDEL be used?

  • aunderwoaunderwo Posts: 3Member

    I am experiencing a problem with the ploidy 1 option.
    Having used GATK2 unified genotyper with the params --sample_ploidy 1 --genotype_likelihoods_model BOTH -rf BadCigar
    I get the following line in a vcf file (see sample in bold)

    Staphylococcus 1553115 . A G 24454.01 . AC=13;AF=0.813;AN=16;BaseQRankSum=1.072;DP=1040;Dels=0.00;FS=32.822;HaplotypeScore=3.3463;MLEAC=13;MLEAF=0.813;MQ=40.20;MQ0=47;MQRankSum=-10.543;QD=32.13;ReadPosRankSum=-1.148;SB=-9.076e+03 GT:AD:DP:GQ:MLPSAC:MLPSAF:PL 1:0,29:29:99:1:1.00:1015,0 1:0,62:62:99:1:1.00:2053,0 1:0,106:106:99:1:1.00:3210,0 1:0,102:102:99:1:1.00:3305,0 1:0,88:88:99:1:1.00:2750,0 1:0,41:41:99:1:1.00:1324,0 1:0,76:76:99:1:1.00:2448,0 1:0,39:39:99:1:1.00:1303,0 0:64,40:104:99:0:0.00:0,1334 1:0,41:41:99:1:1.00:1373,0 1:0,49:49:99:1:1.00:1668,0 0:72,50:122:99:0:0.00:0,1258 1:0,59:59:99:1:1.00:1852,0 1:0,38:38:99:1:1.00:1192,0 1:0,31:31:99:1:1.00:961,0 0:53,0:53:99:0:0.00:0,1633

    The sample in bold is called as WT (genotype 0) with a high GQ despite there being 72 reads of genotype 0 and 50 of genotype 1. Examining the bam file suggests that this is a mapping error in a repetitive phage region

    If I set ploidy to be 2 the equivalent line in the resulting vcf file is

    Staphylococcus 1553115 . A G 24788.02 . AC=28;AF=0.875;AN=32;BaseQRankSum=0.947;DP=1040;Dels=0.00;FS=32.822;HaplotypeScore=3.3463;InbreedingCoeff=0.4286;MLEAC=28;MLEAF=0.875;MQ=40.20;MQ0=47;MQRankSum=-10.096;QD=25.11;ReadPosRankSum=-1.177;SB=-9.871e+03 GT:AD:DP:GQ:PL 1/1:0,29:29:81:986,81,0 1/1:0,62:62:99:1895,156,0 1/1:0,106:106:99:2992,247,0 1/1:0,102:102:99:3169,268,0 1/1:0,88:88:99:2452,193,0 1/1:0,41:41:99:1243,99,0 1/1:0,76:76:99:2283,193,0 1/1:0,39:39:99:1233,105,0 0/1:64,40:104:99:886,0,1706 1/1:0,41:41:99:1298,108,0 1/1:0,49:49:99:1581,129,0 0/1:72,50:122:99:1235,0,2126 1/1:0,59:59:99:1649,132,0 1/1:0,38:38:87:1065,87,0 1/1:0,31:31:69:821,69,0 0/0:53,0:53:99:0,138,1588

    As can be seen from the bold text, the same position is called as heterozygote which based on the number of the reads mapping would be likley except for the fact this is a bacterial haploid genome. Previously I would have discarded this since the heterozygous call indicates mis-mapping as the bam file confirms. I had been hoping to use the sample_polidy option set to 1 for bacterial genomes but this results concerns me. I could obviously filter based on AD but the wonder why the sample was given a high GQ when the polidy is set to 1 and the AD suggests the call of genotype 0 should be doubted.

    Any suggestions on what is going on here?? Many thanks

    Anthony

  • delangeldelangel Posts: 71GATK Dev mod

    The code is actually doing what it's designed to do - when you're using -ploidy 1, there are only 2 possible genotype assignments, and the assignment "0" is by far the most likely one even if 40% of your reads have another base. In the default diploid case, the most likely genotype is the 0/1 one, which is exactly what you're getting.
    Even in the haploid case, there's considerable evidence that favors the "0" genotype (plus the population prior), so you'll get a high value of GQ anyway - your PL values of 0,1258 indicate that, statistically, it's 10^125 likelier that your data came from a reference site than from an alt site based on all the available reads.

  • flowflow Posts: 14Member
    edited January 2013

    In the current documentation (v2.3-9) for the Unified Genotyper there is a caveat stating "We only handle diploid genotypes". Has something changed or can -ploidy still be safely set to 1?

    Post edited by flow on
  • Geraldine_VdAuweraGeraldine_VdAuwera Posts: 8,019Administrator, GATK Dev admin

    That's an old caveat, I'll remove it from the doc. You're good to go.

    Geraldine Van der Auwera, PhD

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