Hello, I see Shaun Jackmans' (abyss-)sam2afg script is available within the Amos source repository. I find this very useful for coercing output from assemblers which which don't track read location into amos by remapping reads against the assembly and converting to afg - not ideal but better than nothing when you don't have the 'real' read locations to work with. This fails, however when reads are aligned with 'bwa mem', which can output multi-part alignments. sam2afg checks for reuse of the same read id (presumably to prevent the generation of non-unique eid values), consequently encountering multiple alignments for a read causes it to die. The following one-line patch allows sam2afg to skip these secondary alignments present in 'bwa mem' output, provided bwa mem has been run with the '-M' argument which sets the SAM 'secondary alignment' flag on the alignments in question. Hopefully this will also be of use to others... Best Regards, James -- Dr. James Abbott Lead Bioinformatician Bioinformatics Support Service Imperial College, London --- abyss-1.3.6/bin/abyss-samtoafg.ori 2015-05-23 23:43:46.797747928 +0200 +++ abyss-1.3.6/bin/abyss-samtoafg 2015-05-23 23:44:09.227747743 +0200 @@ -105,6 +105,7 @@ die unless defined $qqual; $tstart--; # convert to zero-based coordinate + next if $flag & 0x100; # secondary alignment $qid .= "/1" if $flag & 0x40; #FREAD1 $qid .= "/2" if $flag & 0x80; #FREAD2