I am using VarScan v2.4.6, I am failed to output vcf format file when call variant by pileup2snp, my code is: samtools mpileup -f $reference $input.bam | varscan pileup2snp --output-vcf 1 > $output.vcf The result is just the same as without --output-vcf 1, it's do not in vcf format. Could someone help to figure out it, thanks.