GSM9916470: WT, Clone3; Homo sapiens; OTHER
30 million viable cells per genotype were collected (800g, 5min, room temperature), washed with PBS, and recollected. Cells were resuspended in Pierce DSG cross linker (Thermo Fisher #20593 ) at 1x10^6 cells/mL and rotated for 35 minutes at room temperature. Formaldehyde was added dropwise to a final concentration of 1%, and cells were fixed for 10 minutes rotating at room temperature. Tris pH 7.5 was again added dropwise to a final concentration of 0.375M, and cells were quenched for 5 minutes rotating at room temperature. The cells were then collected, washed with cold PBS twice, and aliquoted into 5 million cell pellets. The pellets were snap frozen in liquid nitrogen and stored at -80 °C for Micro-C. Library preparation was performed with the NEBNext Ultra II kit (New England BioLabs #E7645).