GSM5448610: Ly6c+ c-kit+ cells_rep3; Mus musculus; ATAC-seq
Cells were isolated by rapid mechanical disaggregation of TDLNs from established (day 14) untreated B16F10 tumors.  LNs were pooled and stained for CD11c, Ly6c and c-kit (CD117), then the CD11cNEGLy6c+c-kit+ precursor population was sorted and used for ATAC-seq. For each sample, 50,000 nuclei prepared from FACS-sorted Ly6c+c‑kit+ precursor cells 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).