RNA-Seg of Ruditapes philippinarum: female gonad
PolyA-tailed mRNAs were enriched using Oligo(dT) magnetic beads. First-strand cDNA synthesis was initiated from fragmented mRNAs using random oligonucleotides as primers in an M-MuLV reverse transcriptase system. RNA strands were degraded with RNaseH, followed by second-strand cDNA synthesis using dNTPs and a DNA polymerase I system. Purified double-stranded cDNAs were end-repaired, A-tailed, and ligated to sequencing adapters. cDNAs of 250-303 bp were selected, PCR amplified, and purified using AMPure XP beads to create sequencing libraries.