RNA-Seq of mus musculus: adult male liver
mRNA with polyA tail is enriched by Oligo(dT) magnetic beads, and then the obtained mRNA is randomly broken in NEB Fragmentation Buffer with divalent cations. Using the fragmented mRNA as a template, random oligonucleotides as primers, cDNA first strand is synthesized in M-MuLV reverse transcriptase system, then RNA strand is degraded by RNaseH and cDNA second strand is synthesized in DNA polymerase I system with dNTPs as raw materials. The purified double-stranded cDNA is end-repaired, A-tailed and ligated with sequencing adapters, and AMPure XP beads are used to select cDNA of about 200bp, PCR amplification and AMPure XP beads purification of PCR products are performed to obtain the library.