GSM7890165: CD70+, within 1 week of aGvHD onset, 953-P_atac; Homo sapiens; ATAC-seq
Maximum of 50,000 cells were sorted on a cell sorter (FACSAria, BD) and pelleted by centrifugation at 500 × g and 4°C for 10 min. The cell pellet was washed once with cold PBS and centrifuged again. Pellets were resuspended in a 50-μl reaction cocktail containing 1 μl of 0.5% Digitonin (Promega, Madison, WI, USA), 2.5 μl of Tn5 transposase and 25 μl of TD buffer (Nextera DNA library preparation kit, Illumina, San Diego, CA, USA) and incubated for 30 min at 37°C with gentle shaking The tagmented DNA was then purified using Minielute PCR Purification kit (Qiagen, Hilden, Germany) and eluted into 11 μl of elution buffer. The transposed DNA was then amplified using 2.5 μl of indexing primers and 25 μl of Nextera PCR master mix