GSM8155725: Ctrl#2-scATAC; Mus musculus; ATAC-seq
After harvesting bone marrow from the femur and tibia of both legs, the bone marrow was filtered through a 70μm filter. The GMPs were sorted from the bone marrow (CD34+ CD16/32+) by flow cytometry. scATAC-Seq library preparation, cells were initially resuspended in 100 μl lysis buffer (10 mM Tris-HCl, pH7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% Tween-20, 0.1 % IGEPAL, 0.01% digitonin and 1% BSA) and incubated on ice for 3 min. After adding 1 ml wash buffer (10 mM Tris-HCl, pH7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% Tween-20 and 1% BSA), nuclei were collected by centrifugation (500xg, 5 min, 4C). Isolated nuclei were subjected to scATAC-seq library preparation by 10x genomics' scATAC-seq kit. Briefly, isolated nuclei were treated with Tn5 transposases and followed by droplet formation with beads and polymerase. DNA fragments were amplified by PCR in droplets and purified after droplet breakage.