RNA-Seq of scylla paramamosain ovary
Total RNAs of each ovary from one crab were pooled as a ovary sample respectively. Sample mRNA was isolated from total RNA using Sera-mag Magnetic Oligo(dT) Beads (Illumina, USA) and was fragmented. Fragmented and primed mRNA was used as templates to synthesize first strand cDNA and the second strand cDNA. The synthesized cDNA was performed to end-repair, adenylated 3' ends and ligated adapters. Then ligated cDNA was enriched with 15 cycles of PCR amplification using PCR Primer Cocktail. The library fragments were measured by Quant-iT™ PicoGreen® dsDNA Assay Kit to generate cluster.Finally, the library was sequenced on an Illumina HiSeq 3000 platform using TruSeq SBS Kit v3-HS.