GSM3354755: Sample 11_KBM7 2min rep1; Homo sapiens; OTHER
total RNA was isolated using 500μl TRIzol-LS for each sample according to the manufacturer's protocol (Invitrogen Life Technologies) Sample 1,2,9,10: Prior to treatment or run-on, the permeabilized cells were placed on 37℃ for 3 min for thawing. Then 100μL run-on buffer was add to cells for 30sec with pipetting, and stop run-on by adding 500μL TRIzol-LS. After run-on, the experiment followed the PRO-seq protocol (Mahat D B,2016). Sample 3,4,11,12: Prior to treatment or run-on, the permeabilized cells were placed on 37℃ for 3 min for thawing. Then 100μL run-on buffer was add to cells for 30sec with pipetting then place on 37℃ inhibitor for 1.5min, and stop run-on by adding 500μL TRIzol-LS. After run-on, the experiment followed the PRO-seq protocol (Mahat D B,2016). Sample 5,6,13,14: Prior to treatment or run-on, the permeabilized cells were placed on 37℃ for 3 min for thawing. Then 100μL run-on buffer was add to cells for 30sec with pipetting then place on 37℃ inhibitor for 7.5min, and stop run-on by adding 500μL TRIzol-LS. After run-on, the experiment followed the PRO-seq protocol (Mahat D B,2016). Sample 7,8,15,16: Prior to treatment or run-on, the permeabilized cells were placed on 37℃ for 3 min for thawing. Then 100μL run-on buffer was add to cells for 30sec with pipetting then place on 37℃ inhibitor for 31.5min, and stop run-on by adding 500μL TRIzol-LS. After run-on, the experiment followed the PRO-seq protocol (Mahat D B,2016). Sample 17,19: After 10min treatement, 100μL 2-biotin run-on buffer was add to the permeabilized cells. After 3min, 500μL TRIzol-LS was add to sample to stop rn-on, then the experiment followed the PRO-seq protocol (Mahat D B,2016). Sample 18,20: After 10min treatement, 100μL 2-biotin run-on buffer was add to the permeabilized cells. After 6min, 500μL TRIzol-LS was add to sample to stop rn-on, then the experiment followed the PRO-seq protocol (Mahat D B,2016).