design: RNA was harvested using Trizol. After the sample is tested and qualified, oligo(dT)-coated magnetic beads are used to enrich eukaryotic mRNA. Subsequently, fragmentation buffer is added to randomly fragment the mRNA. Using the mRNA as a template, random hexamers are employed to synthesize the first strand of cDNA. Then, buffer, dNTPs, and DNA polymerase I are added to synthesize the second strand of cDNA. Subsequently, AMPure XP beads are utilized to purify the double-stranded cDNA. The purified double-stranded cDNA undergoes end - repair, adenylation, and ligation of sequencing adapters. Then, AMPure XP beads are used for fragment size selection. Finally, PCR enrichment is carried out to obtain the final cDNA library.