RNA-Seq of Homo sapiens: 46,XY LCL; biotinylated RNA pulldown experiment
Cells were treated with 5EU at a final concentration of 400uM for 0-7 hours. Cells were washed and treated with TRIzol (Thermo Fisher Scientific) to isolate RNA. Biotin was attached to metabolically labeled RNAs, followed by reaction quenching and RNA precipitation. RNA was then incubated with streptavadin beads followed by washing. RNA was eluted and precipated and libraries were prepared using SMART-Seq v4 Ultra Low Input RNA kit.