GSM9579382: Mock_D8_Rep2; Mus musculus; ATAC-seq
Whole bone marrow (BM) cells were incubated for 20 min with a purified rat IgG2a anti-mouse CD16/CD32 antibody (BioLegend) to block Fc receptor-mediated nonspecific binding. Cells were then stained for 30 min at 4°C with APC-conjugated antibodies against lineage markers (CD3, B220, Gr-1, Ter119, NK1.1, SiglecF, and CD11b). The cell suspension was subsequently incubated with anti-APC magnetic beads (BioLegend) for 15 min. Cells were passed through LS columns, and the negative (flow-through) fraction was collected and used for subsequent sorting of LSK cells. For bone marrow monocytes, T cells, CMP/GMP populations, and blood monocytes, cells were incubated with an Fc receptor–blocking antibody followed by staining with fluorochrome-conjugated antibodies against surface markers. Cell sorting was performed using a BD FACSAria™ III cell sorter. Gating strategies are shown in the manuscript. Nuclei were resuspended in 50 µl of transposition reaction mix. Transposition was carried out for 30 min at 37°C. DNA was purified using the MinElute PCR Purification Kit. Purified DNA was amplified and indexed by PCR with 10–12 cycles. Amplified libraries were purified using the MinElute PCR Purification Kit, followed by double AMPure XP purification.