GSM4879350: Kasumi-1, siRR treated replicate 2; Homo sapiens; RNA-Seq
Kasumi-1 cells were lysed using TRI Reagent® (Sigma-Aldrich, St Louis, USA) and RNA was purified according to the manufacturer's instruction. Next, 4-thiouridine-incorporated RNA was biotinylated by labelling with 1 mg/ml Biotin-HPDP (Abcam, Cambridge, UK) for 90 minutes at room temperature. Following chloroform extraction, labelled RNA was separated using magnetic streptavidin beads, beads were washed and RNA was eluted in two rounds of elution with 100 μl 100 mM DTT. Finally, RNA was purified using RNeasy MinElute Cleanup Kit (QIAGEN GmbH, Hilden, Germany) RNA libraries were prepared for sequencing using standard Illumina protocols