T. oceanicus 454 accessory gland
The GS FLX Titanium Sequencing Kit XLR70 (Roche) was used. Libraries were synthesized using random primer synthesis (EMD4Biosciences). The technique utilized EcoR I and Hind III linkers to create priming sites at the 3’ end of single-stranded mRNA transcripts. Double-stranded cDNA was synthesized using these primers, and subsequent digestion with EcoR I and Hind III produced fragments that were then ligated onto customized 454 adapters for the sequencing reaction. The resultant libraries were size-selected using Ampure XP beads (Agencourt) and Sizing Solution (Roche) according to the GS-FLX Titanim cDNA rapid library preparation standard method (Roche). Libraries were not normalized prior to sequencing, so read numbers reflected transcript abundance.
University of St Andrews
T. oceanicus 454 accessory gland