Hi all,

I am trying to detect rare variants on a haploid organism using samtools
mpileup + bcftools call (htslib v1.4).
I have a few variants which I am expecting to see however they are not
called as variant.

When I use the command like this:

samtools mpileup -ugf ref.fa 1.bam | bcftools call --ploidy 1 -Am

I get an output like this:
Chromosome    1473246    .    A    G    24.8295    .
DP=80;VDB=0.44194;SGB=-0.693021;RPB=0.139427;MQB=1;MQSB=1;BQB=0.993938;MQ0F=0;AC=0;AN=1;DP4=28,23,12,15;MQ=60
GT:PL    0:255,255

>From "DP4" I can see that there is quite a good 0.65:0.35 ratio for ref and
alt alleles.

Now when I remove the -A flag and add the -v flag  form bcftools call like
so:
samtools mpileup -ugf ref.fa 1.bam | bcftools call --ploidy 1 -vm

I do not get this variant. So my question: Is samtools not able to call
mixed variants?
I thought the -m flag in bcftools was used for this purpose?
If I use --ploidy ==1 does it always pick the major allele?

Any help would be appreciated,
Thanks,
Jody
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