Page: bbmap on pacbio
gravatar for colindaven
3.6 years ago by
Hannover Medical School
colindaven2.6k wrote:

Details on Pacbio SNP calling using bbmap

See C: SNPs Calling Tools

Uncorrected reads

Using non-corrected reads, I could call SNPs using these parameters on 4 human mito sample with coverage of 2000+ in=in.bam out=out_SNV.vcf ref=/lager2/rcug/seqres/HS/mito.fa minquality=1.0 minqualitymax=2 overwrite=t ploidy=1 minscore=2.0 minpairingrate=0 usepairing=f useidentity=f

Note the score parameters are set exceedingly low. I did have them higher, but continually reduced them to get any results at all.

BBMap VCF Qual score distributions of 2-6 were output.

Corrected reads

I called SNPs again with the same parameters on reads that had been corrected by Canu with default settings, genome size set to 16.5k. Corrected coverage was low, around 30 (previously around 2000+). A surprising reduction.

BBMap VCF Qual score distributions of 26-42 were output.

Thanks for the nice VCF formatted pacbio SNPs Brian, now I can process downstream with snpeff etc. This wasn't the case with samtools, Freebayes or SMRT analysis.

snp page • 1.5k views
ADD COMMENTlink written 3.6 years ago by colindaven2.6k

Thanks for the feedback! I'll run some testing on my own and see if I can add a useful set of defaults for raw PacBio reads.

ADD REPLYlink written 3.6 years ago by Brian Bushnell17k

Hi Brian, I am very interested in BBMap' variants calling fuction.I want to use it with pacbio data. But I have not found any artical about the BBMap,especially the variants calling fuction.Could you give my help?

ADD REPLYlink written 3.3 years ago by zhouyanghappy19890

Hello!Could I say that besides the exceedingly low parameters (minquality=1.0 minqualitymax=2 overwrite=t ploidy=1 minscore=2.0 minpairingrate=0) there will be none results ?May I know if you change the software to call variants whith pacbio data at last?

ADD REPLYlink written 3.3 years ago by zhouyanghappy19890
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