title: GSM5471288: m-B6-beta-Ran-1-eHiC; Mus musculus; Hi-C
library_construction_protocol: Mouse beta cells were isolated from C57BL/6 islets and subjected to easy Hi-C (eHi-C) to profile the chromatin loopings. Briefly, 30-40K sorted mouse beta cells were used to generate one eHi-C library. The cells were first lysed with a cell lysis buffer, containing 10mM Tris-HCl pH 7.5, 10mM NaCl, 0.2% NP-40 and protease inhibitor cocktail. The exraced nuclei were then digested with HindIII (NEB, Cat#R3104M). After digestion, the proxity ligation was done in intact inclei with T4DNA ligase (Invitrogen,Cat#15224-025) fot at least seven hours. The DNA were then extracted after reverselinking with proteinase K for overnight and then sujected to DpnII (NEB, Cat#R0543L) digestion. The resulting DNA was then self-ligated with T4 DNA ligase to form self-circles. The DNA species those were not able to self-circled were removed by exonulcease treatment. The self-circles were then linearized with a secondary HindIII digestion. And the resulting DNA were used to generate Libraries following a Illumina Truseq system with modified adapters. Overall, we generated 220M reads for each sample, including 40M Cis-contacts. And conbined together, we generated 246M cis-contact reads. The libraries were constructed following standard Illumina Truseq library preparation protocol.