H1395 cell line RNA sequencing
1 g total RNA was used for following library preparation. The rRNA was depleted from total RNA using single-stranded DNA probe. The poly(A) mRNA isolation was performed using RNA clean beads. The mRNA fragmentation was performed using divalent cations and high temperature. Priming wasperformed using Random Primers. First strand cDNA and the second-strand cDNA were synthesized. The purified double-stranded cDNA was then treated to repair both ends and add a dA-tailing in one reaction, followed by a T-A ligation to add adaptors to both ends. Size selection of Adaptor-ligated DNA was thenperformed using DNA Clean Beads. Each sample was then amplified by PCR using P5 and P7 primers and the PCR products were validated.