title: GSM6432878: single-nucleus Hi-C on mouse ESC - cell 8; Mus musculus; Hi-C
library_construction_protocol: F123 mESCs and NPCs were harvested by accutase treatment and fixed with 1% (v/v) methanol-free formaldehyde at room temperature for 10 minutes. scHi-C libraries were prepared using methods as previously described with slight modifications. In brief, 1-3 million crosslinked mESCs or NPCs were incubated overnight at 37 °C with 200 U MboI followed by proximity ligation at room temperature with slow rotation for 4 hours. Then the nuclei were stained with Hoechst and the single 2N nuclei were sorted by FACS into wells of 96-well plate. After overnight reverse crosslinking at 65 °C, the 3C-ligated DNA in each cell was amplified using GenomiPhi v2 DNA amplification kit (GE Healthcare) for 4-4.5 hours. After purification with AMPure XP magnetic beads and quantification, 10ng WGA product was used to construct library with Tn5. single-nucleus Hi-C