GSM9380657: Naive_BM_sorted_1; Mus musculus; ATAC-seq
Live cells were enriched using the EasySep Dead Cell Removal kit (Stemcell Technologies). Equal numbers of cells from three individual mice under each condition were mixed and the pooled samples were used for downstream sequencing. Bone marrow cells were stained with anti-CD11b and anti-Ly6C antibodies and sorted for CD11b+Ly6Chi monocytes by Bigfoot Spectral Cell Sorter. The purity of sorted monocytes was greater than 98%. For each sample, 50,000 nuclei prepared from FACS-sorted CD11b+Ly6Chi were incubated with 2.5 mL Nextera Tn5 transposase (Illumina) in 50mL 1X transposition reaction mixture in a thermocycler at 37 °C for 1hr. The transposition reaction mixtures were purified with a DNA Clean and Concentrator kit (Zymo Research), and amplified for 11-13 cycles using NEBNext High-Fidelity 2X Master Mix (New England Biolabs) and Nextera Index primers (Illumina). The amplified libraries were size-selected by two rounds of AMPure beads purification (0.5X and 1.2X).