GSM8655706: 1x IFN-g 24h,replicate3,bulk_ATAC-seq; Mus musculus; ATAC-seq
For each condition, 50,000 primary OPCs were collected, washed with PBS, and lysed with 50 μL of lysis buffer (10 mM Tris-HCl pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% IGEPAL (CA-630), 0.1% Tween-20, 0.01% Digitonin) on ice for 3 minutes. After lysis, 1 mL of wash buffer (10 mM Tris-HCl pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% Tween-20) was added, and the nuclei were pelleted by centrifugation at 500g for 10 minutes at 4°C. After aspirating the supernatant, the nuclei were resuspended in 50 μL of transposition mix (25 μL 2x TD buffer (Tagment DNA Buffer), 16.5 μL PBS, 0.5 μL 1% Digitonin, 0.5 μL 10% Tween-20, 2.5 μL Tn5 Transposase (final 100 nM), and 5 μL nuclease-free H2O) and incubated for 30 minutes at 37°C in a thermomixer with 1000 RPM mixing. The DNA was purified both pre- and post-PCR using the Zymo DNA Clean and Concentrator-5 kit (Zymo Research). Libraries were sequenced on an Illumina NovaSeq X with a 51-8-8-50 read setup.