GSM8197318: H358 cells, oeCtrl, Rep1; Homo sapiens; RNA-Seq
Total RNAs were purified with RNeasy Mini Kit (QIAGEN) or RNAprep Pure Cell/Bacteria Kit (TIANGEN). Approximately 5 μg of total RNA was used to deplete rRNA using Ribo-Zero Gold rRNA Removal Kit (Illumina). rRNA-depleted RNAs were fragmented using NEBNext® Magnesium RNA Fragmentation Module (NEB) and then reverse-transcribed by SuperScript™ II Reverse Transcriptase (Thermo Fisher Scientific). Next, U-labeled second-stranded DNAs were synthetized with DNA Polymerase I (E. coli) (NEB), RNase H (NEB) and dUTP Solution (Thermo Fisher Scientific). After adding adapters, the fragments were purified with AMPureXP beads, then treated with Uracil-DNA Glycosylase (UDG; NEB) and were amplified by PCR.