GSM8181667: Donor 3 cortex; Homo sapiens; ATAC-seq
Nuclei were isolated from 2g of kidney cortex or medulla, homogenized in nuclei isolation buffer (250mM sucrose, 10mM Tris pH 7.5, 3mM MgCl2, 0.1% NP40) by gentleMacs using dissociator proram E.01 and filtered through a 200uM filter. Homogenate was resuspended in 1ml of ATAC-Resuspension Buffer with detergents (10mM Tris pH 7.5, 10mM NaCl, 3mM MgCl2, 100ug/mL digitonin, 0.1% Tween, 0.1% NP40) for 3 minutes on ice and then detergents were diluted by resuspending with 14mL ATAC-Resuspension Buffer with tween alone (10mM Tris pH 7.5, 10mM NaCl, 3mM MgCl2, 0.1% Tween). Nuclei were passed through a 40mm filter, counted, and 15,000 nuclei were used for transposition and library generation using the Chromium Next GEM Single Cell ATAC Library & Gel Bead Kit v1.1 (10x Genomics, 1000176). Chromium Next GEM Single Cell ATAC Library & Gel Bead Kit v1.1 with 15,000 nuclei according the manufacturer's protocol.