title: Zebrafish testis
design: mRNA-seq library construction: Total RNA was isolated and quality-checked. Poly(A)+ mRNA was enriched using Oligo(dT) magnetic beads (for eukaryotic samples) or rRNA-depleted using a kit (for prokaryotic samples). mRNA was fragmented with fragmentation buffer. First-strand cDNA was synthesized using random hexamers, followed by second-strand synthesis with DNA polymerase I and RNase H. Double-stranded cDNA was purified with AMPure XP beads. End repair, A-tailing, and adapter ligation were performed sequentially, followed by size selection with AMPure XP beads. PCR amplification was conducted to enrich adapter-ligated fragments, and the final library was purified with AMPure XP beads. Library quality was assessed by Qubit 2.0 (quantification), Agilent 2100 (insert size verification), and Q-PCR (effective concentration determination)