GSM8015474: NIH-3T3 cells, 800 µM 4sU, SLAM-seq, rep B; Mus musculus; OTHER
Cells were harvested using TRI reagent (Sigma) and RNA isolation was conducted using the Zymo DirectZol RNA-microprep kit (R2062) as described by the manufacturer and re-suspended in 1X PBS buffer SLAM-seq was conducted using IAA to mediate U>C conversions at 50oC/20 minutes. The reaction was quenched using excess DTT. RNA was then purified using RNeasy Mini elute kit (Qiagen). TUC chemistry was performed by incubation with OsO4/NH4C for 1h at 40°C. RNA was purified by precipitation. Nucleotide conversion  RNA-seq (SLAM-/TUC-seq)