GSM5261683: 190419Mk3_Ifng; Mus musculus; ATAC-seq
Cells were harvested by TrypLE incubation at 37°C for 5 minutes, collection in cell culture medium and washing in PBS. ATAC-seq was performed as previously described (Buenrostro et al., 2013) with minor adaptations. Primary OPCs were incubated with TrypLE () at 37°C for 5 minutes and collected in cell culture media. 60,000 cells per condition were washed with PBS and lysed with lysis buffer (containing 0.1% IGEPAL (CA-630), 10 mM Tris-HCl pH 7.4, 10 mM NaCl, 3 mM MgCl2) and centrifuged for 20 minutes at 500xg and 4°C. Cells were then resuspended in tagmentation mix (dH2O, 2x TD buffer (Wang et al., 2013) and Tn5 enzyme (Picelli et al., 2014)) for 30 minutes at 37°C. The DNA was purified pre- and post-PCR with the MinElute purification kit (Qiagen) and then PAGE purified to remove adapter dimers. Three replicates per condition were performed with primary OPCs obtained from different litters. Libraries were sequenced on an Illumina Novaseq 6000 with a 50-8-8-50 read set-up.
GEO
GSM5261683