Steady state EU-seq Vehicle__replicate 3
Cells were incubated with 1 mM 5-ethynyluridine (EU) 15 minutes before cell lysis with QIAzol and total RNA isolation with miRNeasy kit (QIAGEN, 217004) with on-column DNAse digestion (QIAGEN, 79256). Nascent RNA was then isolated with the Click-iT Nascent RNA Capture Kit (Thermo, C10365) according to the manufacturer's instruction. EU RNA-containing beads were resuspended in fragment, prime, and elute buffer from the KAPA RNA HyperPrep Kit (Roche, KK8540), heated to 94C for 6 minutes, and further sequencing library preparation was performed following the manufacturers instructions.