GSM7963603
Total RNA was extracted using RNeasy Mini Kit (Qiagen) from frozen cell pellets previously washed with PBS. 1 μg RNA per sample was used as input material. Sequencing libraries were generated at Novogene using NEBNext Ultra RNA Library Prep Kit for Illumina (NEB, USA) following manufacturer's recommendations, index codes were added to each sample. mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X). First strand cDNA was synthesized using random hexamer primer and M-MuLV Reverse Transcriptase (RNase H-). Second strand cDNA synthesis was subsequently performed using DNA Polymerase I and RNase H.