design: We first purified the poly-Acontaining mRNA molecules using polyTattached magnetic beads, fragmented the mRNA into small pieces using divalent cations under elevated temperatures, copied the cleaved RNA fragments into first-strand cDNA using SuperScript II reverse transcriptase (Invitrogen, #18064014) and random primers, and then synthesized second-strand cDNA using DNA Polymerase I, RNase H, and dUTP. Subsequently, these cDNA fragments underwent an end-repair process, a single A base was added, and the adapters were ligated. The products were then purified and enriched with polymerase chain reaction to create the final cDNA library.