GSM1159901: minusDHT_ASO_PRNCR1_hs15l7_3; Homo sapiens; RNA-Seq
Nuclei were isolated from LNCaP cells with appropriate transfection and -/+ DHT treatment. GRO-seq libraries were prepared from 10 million cells based on previously published protocols (Wang et al., 2011). Libraries were sequenced on Illumina Hi-Seq 2000. After harvesting the cells, RNA polymerases were allowed to run-on for 5 minutes at 30C (~100bp) in the presence of sarkosyl and BrUTP. The RNA was then base hydrolyzed and purified. BrUTP-labeled run-on RNA was immunopurified with anti-BrdUTP-coated agarose beads. Run-on RNA was then polyadenylated to generate primer sites for first strand cDNA synthesis. Complementary poly-T primers contain two adaptor sequences separated by an Ape1 restriction site, which after circularization and Ape1 digestion, will terminate both ends, thus permitting cDNA generation, PCR-amplification, and library generation for deep sequencing.
GEO Accession
GSM1159901