GSM9137146: C57BL/6 female 7weeks, Naïve CD8+ T cells, anti-CD3 / anti-CD28 12h, rep2; Mus musculus; ATAC-seq
100,000 cells were washed with PBS and lysed using a cold lysis buffer containing 10 mM Tris-HCl pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% NP-40, 0.1% Tween-20, and 0.01% digitonin. Cells were incubated on ice for 5 minutes. Nuclei were pelleted by centrifugation at 500 x g for 10 minutes at 4°C, washed with 1ml cold lysis buffer without digitonin and Tween-20, and resuspended in transposition reaction buffer. Transposition was carried out in a 50 µL reaction containing 25 µL 2× TD buffer, 2.5 µL Tn5 transposase, 16.5 µL PBS, 0.5 µL 1% digitonin, 0.5 µL 10% Tween-20, and 5 µL nuclease-free water. The reaction was incubated at 37°C for 45 minutes with with tapping every 10 minutes. Directly following transposition, each sample was purified using a DNA clean & ConcentratorTM-5. Following purification, transposed fragments were PCR-amplified using Nextera dual-index primers and NEBNext High-Fidelity Master Mix. PCR products were cleaned using AMPure XP beads.