title: GSM3593913: D4_WT_ATAC; Mus musculus; ATAC-seq
library_construction_protocol: ATAC-seq was performed as described (Corces et al. 2017). In brief, 100,000 cells were collected after once DPBS wash. The cells were resuspended using ice-cold ATAC-RSB buffer (10 mM Tris-HCl, pH7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% NP40, 0.1% Tween-20 and 0.01% Digitonin). After incubation on ice for 3 min, the cells were washed with ice-cold ATAC-RSB buffer without NP40. Then cells were resuspended in transposition mix composed of 0.33×DPBS, 0.01% Tween-20, 1×TD buffer (Nextera DNA Library Prep   Kit, Illumina) and Transposase (100 nM final concentration, Illumina). Transposition reaction was incubated at 37 ºC for 30 minutes in a thermomixer with mixing. Then transposed DNA was purified by MinElute PCR Purification Kit (28004, Qiagen). Eluted DNA was amplified by using NEB Next UltraII Q5 Master Mix for 5 cycles. A real-time qPCR amplification was performed to determine additional cycles. The amplified DNA libraries were size-selected and purified by AMPure beads according to the manufacturer's instructions.
attributes.tag: GEO Accession
attributes.value: GSM3593913