NL-3 (CRX1724835; CRA025785)
Using fragmented mRNA as a template and random oligonucleotides as primers, the first strand of cDNA is synthesized in an M-MuLV reverse transcriptase system. Subsequently, the RNA strand is degraded by RNase H, and the second strand of cDNA is synthesized in a DNA polymerase I system using dNTPs as raw materials. The purified double-stranded cDNA undergoes end repair, A-tailing, and ligation of sequencing adapters. cDNA of approximately 250-300 bp is selected using AMPure XP beads, subjected to PCR amplification, and the PCR products are purified again using AMPure XP beads to finally obtain the library.