FYP-20 (CRX1561153; CRA023833)
(1) Eukaryotic mRNA was enriched with magnetic beads with Oligo (dT); (2) mRNA was randomly interrupted by the addition of Fragmentation Buffer; (3) The first cDNA strand was synthesised using mRNA as a template with random hexamers (six-base random primers), followed by the addition of buffers, dNTPs, RNase H and DNA polymerase I to synthesise the second cDNA strand, and the cDNA was purified using AMPure XP beads; (4) the purified double-stranded cDNA was then subjected to end repair, addition of A-tail and ligation of sequencing junctions, and then fragment size selection was performed using AMPure XP beads; and (5) finally, cDNA library was obtained by enrichment via PCR The cDNA library was then enriched by PCR.