GSM5381370: HBECs, pulse chase Nascent RNA-seq, rep_A_20min; Homo sapiens; RNA-Seq
Cells were trypsinized and washed with PBS. Nuclear fraction was collected with NE-PER nuclear and cytoplasmic extraction reagents (ThermoFisher, 78833). Nuclei pellet was resuspended in Trizol with 1 ml Dounce tissue grinder to homogenize the nuclei. Nascent RNA was purified as described above. Ribosomal RNAs were depleted from the 4sU-selected nascent RNA samples and libraries were prepared with NEBNext RNA library prep kit for Illumina (NEB, E7770).