GSM8286368: INTS11DEG_PRO_1h_rep2; Mus musculus; OTHER
Nuclear run-on experiments were performed on ten million ESC nuclei for three minutes at 30 °C using 25 µM Biotin-11-ATP/UTP/CTP/GTP. After extracting the total RNA, it was fragmented using 0.2 M NaOH for 10 min on ice. Streptavidin beads M-280 were used to purify biotinylated nascent RNAs. 5' end repair was performed using RppH and PNK. SuperScript III was used for reverse transcription after adapter ligation. Following PCR amplification, 140–350 bp libraries were size-selected by AMpure XP and sequenced with single-read runs on an Illumina NextSeq 500.