GSM8951072: IFNAR-/-PBS AMs1; Mus musculus; ATAC-seq
50,000 AMs were washed with cold PBS and resuspend in cold lysis buffer(10 mM Tris-HCl, pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% IGEPAL CA-630). Detail protocol could be find in Curr Protoc Mol Biol. ; 109: 21.29.1–21.29.9. doi:10.1002/0471142727.mb2129s109. Cell lysates were centrifuged at 500g for 10 min at 4℃ and nuclei were resuspended in 50 µL of transposition reaction mix (25 µL of TD buffer + 2.5 µL of Tn5 Transposase + 22.5 µL of nuclease-free water; Illumina, San Diego, CA) and incubated for 30 min at 37 °C. Transposed DNA fragments were purified by using a Qiagen Reaction MiniElute Kit (Qiagen, Hilden, Germany), barcoded with Nextera single indexes (New England Biolabs, Ipswich, MA), and amplified by PCR for 11 cycles using NEBNext High Fidelity 2× PCR Master Mix (New England Biolabs, Ipswich, MA). PCR products were purified using a PCR Purification Kit (Qiagen, Hilden, Germany) and amplified fragment size was verified on an Agilent 2100 Bioanalyzer system (Agilent, Santa Clara, CA). ATAC-seq libraries were quantified by quantitative PCR.