GSM7429053: ATAC_RASGRP1-/-#2; Homo sapiens; ATAC-seq
50,000 cells are sorted in Weill Cornell Medicine Flow Cytometry Core Facility. Then cells were washed with 1000 μl of ice-cold PBS and resuspend the pellet in 25 μl of ice cold 1X ATAC Buffer [20mM Tris-HCl (pH 7.4), 20mM NaCl and 6mM MgCl2]. Incubate for 5 min on ice. Add 25 μl of ice cold ATAC-Detergent-buffer [20mM Tris-HCl (pH 7.4), 20mM NaCl and 6mM MgCl2, 0.2% Igepal CA-630, 0.2% Tween 20 and 0.02% Digitonin]. Mix throughout well. Incubate the samples on ice for another 3 min. Then samples are centrifuged and pellets are collected. Resuspend the pellet in the following transposase mixture (Per reaction): 25 μl 2X TD Buffer (Illumina 15027866), 2.5 μl TDE1 (Illumina 15027865), 16.5 ul PBS, 0.5 ul 1% Digitonin, 0.5 ul, 10% Tween-20 and 5 ul H2O. Incubate the reaction at 37°C for 30 min in thermomixer (Benchmark) set to 500 rpm. Tagmented DNA are purified with Zymo DNA clean and concentrator (Zymo research) according to manufacture instruction. samples are submitted to Weill Cornell Medicine Epigenetics Core facility for library preparation