title: GSM5290949: RNApol _DN63_AAVS1_unstim; Homo sapiens; ChIP-Seq
library_construction_protocol: ATACseq: Approximately 100k CAR-T cells were used from each sample. Nuclei were isolated with ATAC-LB (10mM Tris-HCl pH7.4, 10mM NaCl, 3mM MgCl2, 0.1% IGEPAL) . ChIP: CAR-T cells (3 x 106) were double crosslinked by 50mM DSG (disuccinimidyl glutarate, #C1104 - ProteoChem) for 30 minutes followed by 10 minutes of 1% formaldehyde. Formaldehyde was quenched by the addition of glycine. Nuclei were isolated with ChIP lysis buffer (1% Triton x-100, 0.1% SDS, 150 mM NaCl, 1mM EDTA, and 20 mM Tris, pH 8.0). Nuclei were sheared with Covaris sonicator using the following setup: Fill level – 10, Duty Cycle – 5, PIP – 140, Cycles/Burst – 200, Time – 4 minutes). Sheared chromatin was immunoprecipitated overnight. ATACseq libraries were prepared with Nextera DNA Library Preparation Kit (Illumina). ChIPseq libraries were prepared with Ovation Ultralow Library System V2 (Tecan).