Megastigmus spermotrophus Transcriptome - Larvae sequence
Approximately 10 individual fifth instar larvae were placed into 2 ml Micro tubes (Starstedt) with one volume buffer RLT (Qiagen), 1/100 volume beta-mercaptoethanol and three 3.5mm dia. glass beads (BioSpec Products). Samples were homogenized using the Mini-Beadbeater (BioSpec Products) at half-speed for 90 seconds. The homogenate was centrifuged at 1,300 x g for 3 minutes. Total RNA was extracted using RNeasy (Qiagen), followed by on-column DNase digestion and RNA cleanup, using the manufacture’s guidelines. Next, the RNA extract was purified using an isopropanol precipitation followed by a 100 % ethanol wash and then re-suspended in RNase-free water. The RNA extract was separated on a 1 % agarose gel stained with SYBR Safe (Invitrogen) and visualized under UV light. The RNA quality and quantity was determined using a Nanodrop 2000 instrument (Thermo Scientific) and RNA quality was further analyzed using an Experion Electrophoresis Station (Bio-Rad).    Complimentary DNA library construction with oligo(dT) primers, library fragmentation, size exclusion purifications (target average sequence length of 300 bp) and sequencing on the Illumina sequencing platform (HISeq 2000) were conducted by the BC Cancer Agency Genome Sciences Centre, Vancouver, Canada.