Hi Kanwar, A simple way is to just treat the SAM dataset as a text file and perform some counts similar to those in this tutorial: https://main.g2.bx.psu.edu/u/aun1/p/galaxy101 The basic outline would be to: 1 - remove unmapped with Filter SAM, "The read is unmapped", "no" 2 - convert sam -> interval 3 - group on column 1 (the sequence identifier), then count distinct 4 - select lines with a count of "1" 5 - join back those identifiers with the original sam to extract the alignments Thanks! Jen Galaxy team On 4/25/12 7:38 AM, shamsher jagat wrote:

I have a sam file after running BWASW and want to extract unique (alignments that are aligning once to genome) from this sam file. I read in other posts that I may be able to use Sam tools> filter Sam option to filter the said flag on wise flag. However I could not find whether I have to use default setting of column 2? when I use option of add flags there are different options for pair reads, however my data is single reads. So exactly single read alignments sam file how we extract unique reads. Am I missing something. I can also share history in order to explain my point if required.

Thanks

Kanwar

-- Jennifer Jackson http://galaxyproject.org