title: GSM4059763: PRO-Seq E14 LBH589 6hrs  REP1; Mus musculus; OTHER
library_construction_protocol: Briefly, 30 million nuclei were used for each run-on assay. In vitro nuclear run-on assays were carried out in the presence of ATP, UTP, GTP, and biotin-11-CTP and 0.5% Sarkosyl at 30°C for 3 min. Total nuclear RNA was extracted using Trizol  and ethanol precipitation. RNA was resuspended and hydrolyzed with 0.2 N NaOH, and biotinylated RNA was purified by streptavidin bead binding, which was followed by 3' adaptor ligation, 5' repair, 5' adaptor ligation, reverse transcription, and PCR full-scale amplification. DNA libraries were PAGE purified and submitted for sequencing. Mahat, D. B. et al. Base-pair-resolution genome-wide mapping of active RNA polymerases using precision nuclear run-on (PRO-seq). Nat Protoc 11, 1455-1476, doi:10.1038/nprot.2016.086 (2016).
attributes.tag: GEO Accession
attributes.value: GSM4059763