RNA-Seq of Eriocheir sinensis
The OligoTex mRNA mini kit (Qiagen) was used to isolate poly (A) mRNA after total RNA was collected from the Chinese mitten crab according to the manufacturer's protocol. Fragmentation buffer was added for cutting mRNA into short fragments (100-400 bp). Following the agarose gel electrophoresis (2% TAE-agarose gel), a range of cDNA fragments (200  25 bp) were excised from the gel, and selected for the PCR amplification as templates using the SuperScript Double-Stranded cDNA Synthesis kit (Invitrogen, Camarillo, CA) following the manufacturer's protocol, and PCR Primer PE 2.0 (Illumina, San Diego, CA).