GSM7947389: WT_UT_2; Homo sapiens; ATAC-seq
50,000 viable cells were pelleted and washed in PBS. Subsequently, nuclei were isolated and resuspended in 2x transposition buffer (20mM Tris-HCl pH 7.6, 3mM MgCl2, 20% Dimethyl Formamide) and the tagmentation reaction was performed by adding 2.5µl of Tagment DNA Enzyme 1 (Illumina). The mixture was incubated for 30 minutes at 37°C with mixing at 1000 rpm. Reaction was stopped by 5M Guanidinium thiocyanate (Sigma-Aldrich) and DNA was further purified using AMPure XP beads. Libraries were amplified in PCR reactions by adding 25 µl of NEBNext High Fidelity 2x Master Mix (NEB), 0.8 µl of 10 µM Custom Nextera PCR Primer 1, and 0.8 µl of 10 µM Custom Nextera PCR Barcode to 25 µl of the transposed DNA. The libraries were purified with AMPure XP beads. The concentration of the library was determined using the Qubit dsDNA HS Assay Kit (Thermo Fisher Scientific).