GSM5862020: LT-HSC young rep 2 [scATAC-seq]; Mus musculus; ATAC-seq
10,000 cells (per sample, an equal number of cells (n=2500) from 4 mice were used) of young and old LT-HSCs were sorted into round-bottom 96 well plate containing 0.04% BSA/PBS. For nuclei isolation, cells were lysed with chilled lysis buffer (10 mM Tris-HCl pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% Tween 20, 0.1%Nonidet P40, 0.01 % Digitonin, 1 % BSA) on ice for 3 min. Cells were washed with wash buffer (10 mM Tris-HCl pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% Tween 20, 1 % BSA). Supernatant was discarded and cells were resuspended in 1X Nuclei Buffer (10x Genomics, PN-2000153/ 2000207). Nuclei suspension was directly subjected to the transposition reaction and used for library preparation.