GSM7813949: EXOSC2 AID ESCs, nascent RNA, EU, 4 hr, rep2; Mus musculus; RNA-Seq
ESCs were labeled with 1 mM 5-EU for 10 minutes and harvested by TRIzol with the addition of RNA spike-ins (5-EU labeled S2 cells) for cell number control. Apply purified RNA to DNase I treatment, fragmentation, ethanol precipitation, and biotinylation. Purify biotinylated RNA by phenol-chloroform, followed by two rounds of biotin-affinity purification using M-280 Streptavidin Dynabeads. Eluted RNA was applied for cDNA synthesis and strand-specific library construction by ssDNA-seq Lib Prep Kit (Abclonal, Cat#: RK20222) following manufacturer's protocols.