GSM6723942: KU812_DMSO_3; Homo sapiens; RNA-Seq
Total RNA was purified by RNeasy Mini Kit (Qiagen, #74104) following the manufacturer's instructions. RNA quality was checked using the TapeStation (Agilent). Samples were prepared using the NEBNext Ultra II Directional RNA Library Prep Kit (New England Biolabs (NEB), # E7760L), NEBNext rRNA Depletion kit v2 (NEB, # E7400X) and NEBNext Multiplex Oligos for Illumina (NEB, # E6440L). About 320 ng of total RNA was ribosomal depleted using the rRNA Depletion module. The rRNA-depleted RNA is then fragmented for 10 min determined by RIN (RNA Integrity Number) of input RNA as per protocol and copied into first-strand cDNA using mixed reverse transcriptase and dUTP. Samples undergo end repair and dA-Tailing step followed by ligation of NEBNext adapters. The products were purified and enriched by PCR to create the final cDNA library. The final libraries were checked for quality and quantity by TapeStation (Agilent) and Qubit (Fisher Scientific).