RNA-Seq of RKO-7-3
In eukaryotes, most mRNA have polyA structure, and magnetic beads with Oligo (DT) are used to capture mRNA with polyA structure. The first chain of cDNA is synthesized in M-MuLV reverse transcriptase system using fragmented mRNA as template and random oligonucleotides as primers. RNA chain is degraded by RNaseH, and the second chain of cDNA is synthesized from dNTPs in DNA polymerase I system. The double-stranded cDNA, was purified and repaired, A tail was added and sequenced splice was connected; the cDNA, around 200bp was screened by AMPure XP beads for PCR amplification, and the PCR product was purified by AMPure XP beads. Finally, the sequencing library was obtained.