GSM2302375: iPSC_OX; Homo sapiens; RNA-Seq
mRNA is enriched by using the oligo(dT) magnetic beads, following to be fragmented into short fragments (about 200 bp). Then the first strand of cDNA is synthesized by using random hexamer-primer and M-MuLV Reverse Transcriptase . Buffer, dNTPs, RNase H and DNA polymerase I are added to synthesize the second strand. The double strand cDNA is purified with AMPure XP beads.  Remaining overhangs of the purified  cDNA are converted into blunt ends via exonuclease/polymerase activities. After adenylation of 3’ ends of DNA fragments, NEBNext Adaptor with hairpin loop structure is ligated to prepare for hybridization. In order to select cDNA fragments of preferentially 150~200 bp in length, the library fragments are purified with AMPure XP system. Finally, the final library is gotten by PCR amplification and purification of PCR products by AMPure XP beads.