GSM7732637: D1063-4_MOLM14_IL3_Rep2; Homo sapiens; ATAC-seq
Cells were treated with inhibitors in the presence or absence of IL-3 for 24 h prior to harvest. Omni ATAC-seq was performed as in Corces et al. Briefly, cells were washed in ATAC resuspension buffer (RSB) (10mM Tris-HCl pH7.5, 10mM NaCl and 3mM MgCl2) and then lysed for 3 minutes on ice in RSB buffer with 0.1% NP-40, 0.1% Tween-20. Then the cells were washed with 1ml of ATAC wash buffer consisting of RSB with 0.1% Tween-20. Then the nuclear pellet was resuspended in ATAC transposition buffer consisting of 25μl TD buffer and a concentration of Tn5 transposase enzyme (Illumina) related to the number of input cells, 16.5 μl PBS, 5 μl water, 0.1% tween-20 and 0.01% digitonin and then incubated on a thermomixer at 37°C for 30 minutes. The transposed DNA was then amplified by PCR amplification up to ¼ of maximum amplification, as assessed by a qPCR side reaction. The library was purified using a QIAquick PCR cleanup kit (QIAGEN) followed by ampure (Beckman Coulter) and analysed on a Next Seq 2000 75 using a NextSeq 500/550 High output kit.