GSM4656227: Kasumi-1_DNAseI_minusKTS; Homo sapiens; DNase-Hypersensitivity
DNase I digestions were performed as previously described (Bert AG et al., MCB, 2007). Briefly cells were permeabilized in DNaseI resuspension buffer (60mM KCl, 10 mM Tris pH 7.4, 15 mM NaCl, 5 mM MgCl2 and 300 mM sucrose) and then DNaseI diluted in dilution buffer (60mM KCl, 0.4% NP40, 15 mM NaCl, 5 mM MgCl2, 10 mM Tris pH 7.4 and 2mM CaCl2 was added and incubated at 22oC for exactly 3 minutes. The digestion was terminated by adding cell lysis buffer (300 mM Sodium Acetate, 10 mM EDTA pH 7.4, 1% SDS and 1 mg/ml proteinase K). DNA was then purified using phenol-chloroform extraction. Optimally digested DNA then underwent library preparation using the KAPA Hyper Prep kit (Roche) and size-selected for 200-300bp fragments. Libraries were sequenced on a NextSeq 550 (Illumina) and run in single-end mode for 75 cycles.