GSM1755013: LP-1_500nM_JQ1_10min_GRO-seq_rep2; Homo sapiens; OTHER
Briefly, nuclei were isolated and nuclear run-on reactions were performed in the presence of ATP, GTP, CTP and BrdU.  RNA was isolated and base hydrolyzed to ~100 nt.  Nascent RNA was isolated using anti-BrdU antibodies conjugated to agarose beads. Briefly, A poly A tail was added to each nascent RNA and used to prime first strand cDNA synthesis that added Illumina adaptors to one end of each nascent RNA.  First strand cDNA was circularized and digested to give linear cDNAs flanked by adaptor sequences.  Linear cDNAs were PCR amplified for 16 cycles. Libraries were size selected (175-400 bp) on a 2% agarose gel.