RNA-seq of Mytilus galloprovincialis: mantle
Total RNA was isolated with the E.Z.N.A. Mollusc RNA Kit (Omega Bio-Tek, USA) according to the manufacturer's instructions. From pooled total RNA, mRNA was isolated with Dynabeads mRNA DIRECT Micro Kit (Ambion). In the first step of the cDNA library preparation, mRNA was fragmented with the RNase III enzyme then purified with the help of magnetic beads and finally quantified with the Qubit Fluorometer (Invitrogen). In the second step, fragmented mRNA was hybridized and ligated with an adaptor mixture and reverse transcribed with Superscript III Enzyme Mix. Before and after the amplification of the cDNA library, cDNA was purified using magnetic beads.