GSM7787328: Kc167,nascentRNA,effkd, bio rep2; Drosophila melanogaster; RNA-Seq
Kc cells were incubated with 0.2 mM Ethylene uridine (EU) for 1 h and RNA was extracted with RNeasy Plus Mini Kit (Qiagen). Samples for nascent RNA-seq (neuRNA-seq) were labeled and captured by using Click-iT Nascent RNA Capture Kit (Thermo Fisher Scientific) according to the manufacturer's protocol. Biotinylation of RNA by Click reaction was performed with 0.5 mM biotin azide using 1 µg of EU-RNA, and then the biotinylated RNA was captured with 50 μL Dynabeads Streptavidin T1 magnetic beads. The nascent EU-RNA libraries were prepared using Universal Plus Total RNA-Seq with Drosophila AnyDeplete (Tecan Genomics) and sequenced on Novaseq using paired-end sequencing at the NHLBI DNA Sequencing and Genomics Core Facility.