RNA-seq of untreated MH7A cells(repeat 2)
Magnetic beads with Oligo(dT) are utilized to enrich mRNA from eukaryotic organisms. Subsequently, a fragmentation buffer is added to randomly fragment the mRNA. Using the mRNA as a template, the first strand of cDNA is synthesized using random hexamers. Following this, buffer, dNTPs, and DNA polymerase I are added to synthesize the second strand of cDNA. Thereafter, AMPure XP beads are employed to purify the resulting double-stranded cDNA. The purified double-stranded cDNA undergoes end repair, adenylation, and ligation of sequencing adapters. Subsequently, AMPure XP beads are used for size selection of the fragments. Ultimately, PCR amplification is conducted to produce the final cDNA library.