GSM5448607: GMP-21dL/14dR [ATAC-seq]; Mus musculus; ATAC-seq
LSK and GMP cells from the BM of LIP-subjected mice or unligated control mice were sorted by flow cytometry. Nuclei isolation for single-cell ATAC sequencing was performed according to the “Nuclei Isolation for Single Cell ATAC Sequencing demonstrated protocol” (10x Genomics). Nuclei suspension was mixed with the tagmentation mix and incubated for 1 hr at 37°C. After mixing with a barcoding mix, the nuclei were loaded into a 10x chip H together with barcoded beads and partitioning oil (Chromium Next GEM Chip H Single Cell Kit v1.1, Chromium Next GEM Single Cell ATAC Library & Gel Bead Kit v1.1, 10x Genomics) and encapsulated using the Chromium controller (10x Genomics). The Gel Bead-In EMulsions (GEMs) was transferred into a PCR tube and amplified for 12 cycles in a thermocycler. The barcoded DNA was purified and subjected to an index PCR for 11 cycles.