GSM9033044: ATAC, control, rep3; Mus musculus; ATAC-seq
1 × 10^5 cells were lysed in 50 µl lysis buffer (10 mM Tris-HCl, pH 7.5; 10 mM NaCl; 3 mM MgCl₂; 0.1% NP-40; 0.1% Tween-20; 0.01% digitonin) on ice for 15 minutes. Subsequently, 1 ml wash buffer (10 mM Tris-HCl, pH 7.5; 10 mM NaCl; 3 mM MgCl₂; 0.1% Tween-20) was added and the samples were gently mixed before centrifugation at 500 g for 10 minutes at 4°C. The pellet was resuspended in 50 µl tagmentation buffer (1× TD buffer; 0.1% Tween-20; 0.01% digitonin; and 2.5 µl Tn5 in PBS) and incubated at 37°C for 30 minutes on a thermomixer at 1000 rpm. Tagmented DNA was purified using the Qiagen MinElute Reaction Cleanup kit and eluted in 10 µl EB. Libraries were amplified with Nextera indexed P5/P7 primers and NEBNext High-Fidelity 2× PCR Master Mix (8–10 cycles), purified by gel extraction (selecting fragments of 150–500 bp), recovered using Qiagen QG buffer, quality controlled by Bioanalyzer, quantified by Qubit HS kit