RNA-Seq of Ovis aries:adult female lung
After enrichment of eukaryotic mRNA with a polyA tail by magnetic beads with Oligo(dT), the mRNA was broken with buffer. The fragmented mRNA was used as a template and random oligonucleotides were used as primers to synthesize the first strand of cDNA in M-MuLV reverse transcriptase system, followed by degradation of RNA strands with RNaseH and synthesis of the second strand of cDNA with dNTPs under DNA polymerase I system. The purified double-stranded cDNA was end-repaired, A-tailed and sequenced, and the cDNA of about 200bp was screened by AMPure XP beads, PCR amplified and the PCR product was purified again by AMPure XP beads to obtain the final library.