GSM8019348: NIH-3T3 cells, methanol fixed cells, no4sU, rep A; Mus musculus; OTHER
Cells subject to 4sU-labeling 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 conversion was performed in tubes or in methanol fixed cells. It was conducted using IAA to mediate U>C conversions at 50oC/30 minutes, conversion in “Tube” samples was done at 37°C/1h. The reaction was quenched using excess DTT. RNA was then purified using RNeasy Mini elute kit (Qiagen)3