GSM5140844: MV411_ATAC_steadystate_REP1_191120_TAAGGCGA_S1; Homo sapiens; ATAC-seq
Five million cells from each cell line (MV4;11 and MOLM13) were washed in PBS and resuspend in a hypotonic buffer (50 mM KCl, 10 mM MgSO4, 5 mM HEPES, 0.05% NP-40, 1 mM PMSF, 3 mM DTT) to generate intact nuclei and washed once in a nuclear wash buffer (10 mM Tris HCl pH 7.5, 10 mM NaCl, 3 mM MgCl2). The resulting nuclei were counted and 50,000 nuclei per cell line were used for the transposition reaction. Nuclei were incubated in 1X TD Buffer (Illumina) and 2.5 µM of Transposase enzyme (Illumina) for 30 minutes at 37 °C. Then the transposed DNA was extracted from nuclei using the MiniElute PCR purification Kit (Qiagen). PCR reactions were carried out to amplify transposed DNA fragments and prep samples for sequencing using NEBNext High-Fidelity PCR kit (NEB) with a universal Nextera primer and a unique indexing primer for each cell line. NEBNext® High-Fidelity PCR kit (NEB) with a universal Nextera primer