RNA-Seq of Holothuria leucospilota: adult female gonad
Total RNA was extracted from gonadal tissues using TRIzol Reagent (Thermo Fisher Scientific, USA, cat No. 11596-026). RNA purity, concentration, and integrity were assessed with an Agilent 5400 Bioanalyzer (Agilent Technologies, USA). Polyadenylated mRNA was enriched via Oligo(dT) magnetic beads, followed by first- and second-strand cDNA synthesis. Fragmented cDNA underwent end repair, adenylation, adapter ligation, size selection, PCR amplification, and purification to construct sequencing libraries.