S03-O
After the RNA sample passed quality control, eukaryotic mRNA was enriched using magnetic beads coated with Oligo(dT). Fragmentation buffer was then added to break the mRNA into short fragments. Using the mRNA as template, first-strand cDNA was synthesized with random hexamers. Subsequently, buffer, dNTPs, DNA polymerase I, and RNase H were added to synthesize the second-strand cDNA. The double-stranded cDNA was purified using AMPure XP beads. The purified double-stranded cDNA underwent end repair, A-tailing, and adapter ligation, followed by size selection with AMPure XP beads. Finally, PCR amplification was performed, and the PCR product was purified with AMPure XP beads to obtain the final sequencing library.