Adult-Club  from Odontodactylus scyllarus
Total RNA was extracted from the club and whole larvae using a bead beating procedure. Frozen samples stored in RNA-later were spun down and RNA-later removed. Trizol and 0.5 mm Zirconia beads were added. The sample was homogenized using a Mini-BeadBeater-1, running at full power at 1 min intervals followed by cooling on ice. This process was repeated for 5 cycles. The resulting homogenate was the passed through a QIAshredder (Qiagen) to remove viscosity and RNA was purified using the RNeasy Mini RNA isolation kit. Transcriptomic libraries were prepared using the ScriptSeq RNA preparation kit (version 2). The pooled libraries were sequenced on a HiSeq 2000 with 2x101 read length. The raw fastq reads were checked with FastQC (http://www.bioinformatics.babraham.ac.uk/projects/fastqc/) and quality trimmed with Trimmomatic (7). The paired end reads were then pooled and a denovo transcript assembly was performed with Trinity (8). To estimate the expression level of each transcript, each library was then analysed individually against this reference using RSEM