No output from Samtools pileup

[pravee1216]

I tried to call consensus using samtools pileup from the filtered BAM file which has only reads with no mate. But the result was empty. The following is what I did:

1) From the complete BAM, I extracted reads (mapQV >8) which have no mate (mate unmapped) using "samtools view -f 0x8" - There are ~35m reads.
2) removed all duplicate reads using Picard
3) provided this filtered file to samtools for consensus calling with options "-c", "-Q 8", "-f ref.fas" ( i tried -m, -M as well)

Question here is 1) Why does pileup fail to call consensus 2) Did i make any mistake or do I modify any parameters 3) Is there any other tool can do this?

Any light to this will be a great help.

Thanks.

Praveen

[ekg]

Some tests I just did show that samtools mpileup only works for proper pairs. Can anyone else confirm this?

I was not able to call any variants until I marked the pairs with bamtools resolve.