Hi, This is already an old thread i know, but i guess it is better to call for new one. I am working on a project that singlets are needed. I have v3.2.1, here is the simplest command i use: mira -project=s1 -fasta -job=denovo,genome,accurate,454 -notraceinfo 454_SETTINGS -AS:mrpc=1 -OUT:sssip=1 i have no output for singlets, but it is important. thank you in advance.. Visam