GSM9242518: Jurkat cell resting ATAC-seq rep2; Homo sapiens; ATAC-seq
Jurkat cells were pretreated with DNase I (Thermo Fisher Scientific, EN0521) to digest DNA from dead cells and to eliminate free-floating DNA. The cells were counted using Crystal Violet (Sigma-Aldrich, C0775), and approximately 50,000 cells from each sample were resuspended in 1 ml of cold ATAC-Resuspension Buffer (RSB), which contained 3 mM MgCl2 (Sigma-Aldrich, M2670), 10 mM Tris-HCl (pH 7.5) (Thermo Fisher Scientific, 15567027), and 10 mM NaCl (Sigma-Aldrich, S7653) in sterile H2O. After centrifugation, the supernatant was discarded, and the cell pellets were resuspended in 50 μl of cold RSB containing 0.1% Tween-20 (Sigma-Aldrich, P1379), 0.1% NP-40 (Thermo Fisher Scientific, FNN0021), and 0.01% Digitonin (Sigma-Aldrich, D141). Following lysis, 1 ml of RSB containing 0.1% Tween-20 was added, and the tube was inverted several times to ensure thorough mixing. After centrifugation, the supernatant was removed, and the nuclei were resuspended in 50 μl of Transposition Mix (Vazyme, TD501). The transposition reaction was then incubated at 37°C for 25 minutes with shaking at 1,500 x g. The reaction products were purified using the QIAquick PCR Purification Kit (QIAGEN, 28106). The ATAC-seq library was prepared with the TruePrep DNA Library Prep Kit V2 for Illumina (Vazyme, TD501) and the TruePrep Index Kit V2 for Illumina (Vazyme, TD202), following the kit's instructions.