title: GSM5232933: Donor_1_YY1_B2_ATAC; Homo sapiens; ATAC-seq
library_construction_protocol: We harvested, counted, and treated each T cell culture with 200U/mL of DNase (Worthington Cat # LS002007) for 30 mins at 37 degrees. We then transferred 60,000 cells of each T cell culture into individual wells of a 96-well plate and washed cells once with PBS and once with RSB (10mM Tris HCl pH 7.5, 10mM NaCl, 3mM MgCl2). Cells were lysed in 50uL of cell lysis buffer (0.1% NP40, 0.1% Tween-20, 0.01% Digitonin in RSB) on ice for 3 min. To the cell lysate, we then added 150ul of RSB with 0.1% Tween-20 to each well and pelleted nuclei at 500g for 10 min at 4 degree. Cells were resuspended in 50ul transposition mix (25ul 2X TD buffer, 16.5ul 1X PBS, 0.5ul 10% Tween, 0.5ul 1% Digitonin, 2.5ul Tn5 transposase, and 5ul H2O) and transposition was performed at 37 degrees for 30 min with 300rpm shaking. Transposed fragments were purified using ZR-96 DNA Clean & Concentrator-5 Kit (Zymo D4024) and libraries were generated using PCR amplification with Nextera adapters and purified using Ampure beads. ATAC-Seq libraries were sequenced on a Novaseq with paired end 100 bp reads.