title: ATAC-Seq of monocytes after alcohol use
design: Following the Omni-ATAC protocol, 2x104 purified monocytes 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 minutes (54). Immediately after lysis, nuclei were spun at 500 g for 10 minutes at 4C to remove supernatant. Nuclei were then incubated with Tn5 transposase for 30 minutes at 37C. Tagmented DNA was purified using AMPure XP beads (Beckman Coulter, Brea, CA) and PCR was performed to amplify the library under the following conditions: 72C for 5 min; 98 for 30s; 5 cycles of 98C for 10s, 63C for 30s, and 72C for 1min; hold at 4C. Libraries were then purified with warm AMPure XP beads and quantified on a Bioanalyzer (Agilent Technologies, Santa Clara CA). Libraries were multiplexed and sequenced on a NextSeq (Illumina).