GSM8758032: WT Clone 1, day 35, replicate 1; Homo sapiens; ATAC-seq
Organoids were resuspended and dounced in 300 ul of RSB buffer (10 mM Tris-HCl pH 7.4, 10 mM NaCl, and 3 mM MgCl2 in water) containing 0.1% NP-40, 0.1% Tween-20, and 0.01% digitonin. Homogenates were transferred to a 1.5 ml microfuge tube and incubated on ice for 10 min. Nuclei were filtered through a 40 μm cell strainer and nuclei were washed with RSB buffer and counted. Fifty thousand nuclei were resuspended in 50 μl of transposition mix38 (2.5 μl transposase (100 nM final), 16.5 μl PBS, 0.5 μl 1% digitonin, 0.5 μl 10% Tween-20, and 5 μl water) by pipetting up and down six times. Transposition reactions were incubated at 37 °C for 30 min in a thermomixer with shaking at 1000 r.p.m. Reactions were cleaned with Qiagen columns. standard ATAC-seq library construction