GSM9181194: PRO-seq, THP1, OTS 4 hours, release with AZ5576 10 min; Homo sapiens; OTHER
For PRO-seq nuclei were isolated and subjected to in vitro transcription reaction (37C, 3min) with 0.05mM biotin-11-CTP. Nuclear RNA was isolated by phenol-chloroform extraction and ethanol precipitation. Nuclear RNA was hydrolysed using 1M NaOH and buffer exchange was performed using BioRad Micro Bio-Spin P30 columns. Biotin-11-CTP labelled RNA was enriched using Streptavidin-linked dynabeads and enriched RNA was isolated by phenol-chloroform extraction and ethanol precipitation. Libraries were generated using the NEBNext Ultra II Directional RNA Library prep kit and NEBNext rRNA Depletion kit as per manufacturer's instructions