GSM9105246: Transcriptome-wide SHAPE-MaP analysis of DMSO-treated cells, 2A3 probed, Replicate 1; Homo sapiens; RNA-Seq
RNA samples in TRIzol™ were mixed with 0.2 volumes chloroform (cat. 319988, Merck) by vigorous vortexing for 15 sec, after which samples were allowed to sit at room temperature for 2 min, before centrifugation at 12,500g for 15 min (4°C). After phase separation, the upper aqueous phase was transferred to a clean 2 mL tube and mixed with 1 volume 2-propanol (cat. 190764, Merck) by vigorous vortexing for 15 sec. Samples were allowed to sit at room temperature for 10 min, before centrifugation at 17,000g for 10 min (4°C). Supernatant was discarded and the RNA pellet was washed twice in 75% ethanol. The RNA pellet was then allowed to air-dry for 10 min, before resuspension in nuclease-free water, pre-warmed at 70°C. Residual gDNA was removed by digestion with TURBO DNase I at 37°C for 30 min.