Molecular phylogeny and evolution of the proteins encoded by coleoid (cuttlefish, octopus, squid) posterior venom glands
Total RNA extracted using the standard TRIzol Plus method (Invitrogen). Extracts were enriched for mRNA using standard RNeasy mRNA mini kit (Qiagen) protocol. mRNA was reverse transcribed, fragmented and ligated to a unique 10*base multiplex identifier (MID) tag prepared using standard protocols and applied to one PicoTitrePlate (PTP) for simultaneous amplification and sequencing on a Roche 454 GS FLX  Titanium platform (Australian Genome Research Facility). 50,000 sequences for each sample were read. Automated grouping and analysis of sample-specific MID reads informatically separated sequences from the other transcriptomes on the plates, which were then post-processed to remove low quality sequences before de novo assembly into contiguous sequences using v 3.4.0.1 of the MIRA software program.