GSM8194091: ADSC_Control; Mus musculus; Hi-C
About 1 million cells were pelleted and cross-linked with 1% methanol-free formaldehyde for 10min at room temperature, and quenched with 125 mM glycine. Then the cells were treated with lysis buffer and cell nuclei were pelleted by centrifuging. The nuclei were then incubated with restriction enzyme MseI. After enzyme digestion, a biotin-linker was added to the chromatin fragment, followed by fragment-end phosphorylation and in situ proximity ligation. After ligation, reversal of cross-linking was performed by adding proteinase K and 10% SDS to the reaction product, and DNA was extracted using phenol:chloroform:isoamyl alcohol (25:24:1). In situ DLO Hi-C libraries were constructed as previously described: “Lin, D. et al. Digestion-ligation-only Hi-C is an efficient and cost-effective method for chromosome conformation capture. Nature genetics 50, 754 (2018).”