GSM4159472: GSK3_KO_ESC_SL_GRO-seq_rep1; Mus musculus; OTHER
Nuclei from ESCs were extracted, and run-on-transcribed with BrUTP (Sigma, B7166) and other NTPs at 30 °C for 5 min. Nascent RNA was enriched by agarose coated anti-BrUTP (Sant Cruz, sc-32323). Poly(A) tail was added to the nascent RNA by Poly(A) Polymerase (NEB, M0276S) to synthese cDNA with oligo(dT) primer. GRO-seq libraries were amplified by PCR for 10 cycles and seperated by 10% TBE gel. The bands ranging from 160 bp to 300 bp were cutted and purified by isopropanol precipitation. Libraries were sequenced on a NovaSeq 6000  sequencer according to the manufacturer's instructions with 12Gbase PE150 reads.
GEO Accession
GSM4159472