RNA-seq of Rattus norvegicus lung
After the sample test is qualified, the library is constructed. The main process is as follows:(1) Enrichment of eukaryotic mRNA with magnetic beads with Oligo (dT);(2) mRNA was randomly interrupted by Fragmentation Buffer.(3) mRNA was used as template to synthesize the first cDNA strand with six-base random hexamers, and then the second cDNA strand was synthesized by adding buffer, dNTPs, RNase H and DNA polymerase I. cDNA was purified by AMPure XP beads.(4) The purified double-stranded cDNA was then end-repaired, A-tail was added and sequencing joints were connected, and AMPure XP beads were used for fragment size selection;(5) cDNA library was obtained by PCR enrichment.