GSM9005479: HAP1 cells, untreated control, biol rep 1; Homo sapiens; RNA-Seq
RNA was isolated using Trizol-chloroform extraction. Briefly, 800 μL Trizol was added directly to the wells, incubated for 5 minutes, and then transferred to DNase/RNase-free 1.5 mL eppendorf tubes. After adding 160 μL chloroform, samples were shaken vigorously for 15 seconds and incubated at room temperature for 3 minutes. Phase separation was achieved by centrifugation at 12,000×g for 15 minutes at 4 °C, and then the aqueous phase was transferred to a new tube. Next, 400 μL isopropanol was added and then samples were incubated for 10 minutes at room temperature. RNA was then pelleted by centrifugation at 12,000×g for 10 minutes at 4 °C. The RNA pellet was washed with 800 μL 75% ethanol, vortexed briefly, and centrifuged at 7,500×g for 5 minutes at 4 °C. The supernatant was removed, and pellets were spun again for 5 minutes. Residual ethanol was removed and pellets were air-dried at room temperature for 5–10 minutes until the pellet turned translucent. RNA was resuspended in 40 μL DNase/RNase-free water, incubated at room temperature for 10 minutes, and gently mixed. Performed by Novogene using a paired-end 150 bp sequencing strategy