ATAC-Seq from purified Splenic Macrophages from controls
100000 purified splenic monocytes were lysed in lysis buffer (10mM Tris-HCl (pH 7.4), 10 mM NaCl, 3 mM MgCl2, and NP-40 for 10 min on ice to prepare the nuclei. Immediately after lysis, nuclei were spun at 500g for 5 min to remove the supernatant. Nuclei were then incubated with Tn5 transposase and tagmentation buffer at 37C for 30 min. Stop buffer was then added directly into the reaction to end the tagmentation. PCR was performed to amplify the library for 15 cycles using the following PCR conditions: 72C for 3 min; 98C for 30s and thermocycling at 98C for 15s, 60C for 30s and 72C for 3 min; following by 72C 5 min. Libraries were then purified with AMPure (Beckman Coulter, Brea CA) beads and quantified on the Bioanalyzer (Agilent Technologies, Santa Clara CA).