GSM8031747: TT-seq-NELFCAID-DMSO4h-NVP2_30m-Rep2; Homo sapiens; OTHER
Nascent RNA was labeled with 500 µM 4sU for 10 min in the cell culture. Total RNA was extracted using TRIzol. RNA was fragmented at 94°C for 2 min usng NEBNext Magnesium RNA Fragmentation Module. MTSEA-biotin-XX was used for biotinylation of fragmented 4sU RNA. The labeled RNA was purified using Dynabeads MyOne Streptavidin C1 and eluted by DTT. rRNA was depleted using NEBNext rRNA Depletion Kit. The remaining labeled RNA was used for library preparation using NEBNext Ultra II Directional RNA Library Prep Kit for Illumina.