GSM4041006: Hi-C Scc1 flfl NSN FGO 4; Mus musculus; Hi-C
Fully grown denuded GV (germinal vesicle) oocytes were isolated from 2-3 month old Scc1 flox/flox and Scc1 flox/flox;Zp3-Cre female mice by puncturing of ovaries with hypodermic needles. Oocytes were incubated for at least 2 hours before the procedure to ensure viability. Before formaldehyde fixation, the zona pellucida was removed by treatment with Acidic Tyrode's solution (Sigma). Oocytes were fixed in with 2% formaldehyde for 15 minutes at room temperature. Oocytes were washed through drops of complete M2. For SN and NSN distinction oocytes were stained for 15 min with 0.2 μg/ml Hoechst (33342, Invitrogen). Cells were then imaged using a confocal microscope (LSM510 Axiovert 200M, Zeiss) and sorted according to their chromatin structure before proceeding with the Hi-C protocol. Single-nucleus Hi-C libraries were produced as described (Flyamer et al., 2017).