GSM9474401: CD4 T cell donor 3 replicate 2 ATAC; Homo sapiens; ATAC-seq
150,000 cells were harvested, centrifuged at 500g for 5 minutes before being resuspended in ATAC lysis buffer (10 mM Tris HCL, pH 7.5, 10 nM NaCl, 3mM MgCl2, 0.1 % Igepal CA-630) and immediately span at 40C for 10 minutes at 500g. the resulting nuclear pellets were PBS washed before being resuspended in a tagmentation buffer (20 mM Tris-HCl pH 7.5, 10mM MgCl2, 20 % Dimethyl Formamide) with Tn5 transposase (Illumina, 20034197). This reaction was incubated for 30 minutes at 370C. The reaction was quenched was buffer PB of the Qiagen MinElute DNA purification kit (Qiagen 28004) and DNA was purified following the manufacturer's instructions. The purified DNA was then amplified using the Q5 high-fidelity mastermix (NEB M0491S) with Illumina Nextera Sequencing primers. Capture enrihcment was performed using the Roche Hypercapture kit and reagents.