How to call variant minimum 3 read coverage to make consensus?

How to call variant minimum 3 read coverage to make consensus?

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I have a query regarding consensus sequence assembly where reference bases are replaced with variants with a minimum of 3 read depths, using bcftools using the below command.

bcftools mpileup -f ref.fasta mapped.bam | bcftools call -c | vcfutils.pl vcf2fq -d 3 > cons.fq

Still, if I check the consensus sequence in IGV the variant is not in the consensus sequence at low coverage site.

Even I mapped reads in CDS sequence using “end to end ” alignment so I already prohibit soft clipping.

I would like to compare pairwise sequence distance further so every variant is important even if ts low coverage side with a minimum of 3 reads depth.

Is there any parameter I am missing in my above command?

Kindly guide me!
Thank you


bcftools


mpileup

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