GSM4633456: ATAC_H1 WT mESCs, replicate 1; Mus musculus; ATAC-seq
Cells were harvested using trypsin and counted. 50000 cells were centrifuged at 300g for 5 minutes, followed by a wash in PBS, and were centrifuged again at 300g for 5 minutes. Cells then were lysed in 50 μl of lysis buffer (10mM Tris-HCl pH7.4, 10 mM NaCl, 3 mM MgCl2, and 0.1% IGEPAL CA) while being centrifuged at 250g for 10 minutes in 4 °C. The pellet was resuspended in a 50 μl of transposition reaction (25 μl of X2 TD Buffer (Illumina), 2.5 μl transposase and 22.5 μl of nuclease-free water). The reaction was performed at 37 °C for 30 minutes. Immediately after, the DNA products were purified using a MiniElute kit (Qiagen). Purified transposed DNA was amplified and barcoded using custom Nextera PCR primers 1 and 2, in a 12 cycle PCR reaction in KAPA HiFi HotStart (Roche). The libraries were purified using a MiniElute kit (Qiagen) and sequenced using Illumina NGS.
GEO Accession
GSM4633456