GSM7653909: WT_Cl13_2 [ATAC-seq]; Mus musculus; ATAC-seq
Following the Omni-ATAC protocol, samples were lysed in lysis buffer (10mM Tris-HCl (pH 7.4), 10mM NaCl, 3mM MgCl2, 10% Np-40, 10% Tween, and 1% Digitonin) on ice for 3 minutes68. Immediately following lysis, nuclei were spun at 500g for 10min at 4°C to remove supernatant. Nuclei were then incubated with Tn5 transposase for 30min at 37°C. Tagmented DNA was purified using AMPure XP beads and PCR was performed to amplify the library under the following conditions: 72°C for 5min; 98°C for 30s; 5 cycles of 98°C for 10s, 63°C for 30s, and 72°C for 1min; hold at 4°C. Libraries were then purified with warm AMPure XP beads and quantified on a Bioanalyzer. Libraries were multiplexed and sequenced to a depth of 50million 100bp paired reads on a NextSeq 500