GSM6625149: core essential_tumor suppressor CHyMErA Library; Homo sapiens; OTHER
Cells were lysed in 12 mL of TEX buffer (10 mM Tris-HCl, pH 7.5; 1 mM EDTA, pH 7.9; 0.5 % SDS) containing ribonuclease A (RNase, 20 mg/mL, Carl Roth GmbH, 7156.2) and proteinase K (20 mg/mL, Carl Roth GmbH, 7528.6) and incubated overnight at 37°C under constant agitation. The next day after complete lysis, 4 mL of 5 M NaCl was added, and the mixture was shaken vigorously and incubated at 4°C for 1 hour. The tube was centrifuged at 15,000 g for 1 hour at 4° C. The supernatant containing the DNA was transferred into a new tube. 24 mL of ice-cold absolute ethanol was added, and mixed and the solution was incubated overnight at -20°C. The next day, the tube was centrifuged at 15,000 g for 1 hour at 4°C. The supernatant was removed and the DNA pellet was washed with 10 mL of ice-cold 70% ethanol and mixed very well. The mixture was centrifuged at 15,000 g for 1 hour at 4°C and the supernatant was removed. Air-dried DNA pellets were resuspended in 1 mL of sterile water. 