GSM5904775
Alveolar macrophages (25,000) were lysed in 25 ml lysis buffer (10 mM Tris-HCl pH7.5, 10mM NaCl, 3mM MgCl2, 0.1% IGEPAL CA-630), followed by addition of 1.25 ml Tagment DNA enzyme I (Illumina 15027865). DNA was extracted with the Zymogen ChIP DNA purification kit (Zymo). DNA was amplified using the Nextera Primer Ad1, Nextera Barcode Primers Ad2.n, SYBR Green, and NEBNext High-Fidelity 2X PCR Master Mix for 5-15 cycles. PCR reactions were size selected for 160-280 bp by gel extraction (10% TBE gels, Life Technologies EC62752BOX) and then 75 bp single-end sequenced for 51 cycles on the Illumina HiSeq 4000 following the manufacturer's protocols.