GSM7508557: UMI-4C in human neural retina, ABCA4 Interacting Region 12 (IR12) as viewpoint, Ret_2_R_ABCA4_UMI_4; Homo sapiens; OTHER
Post-mortem human neural retina and RPE/choroid mixtures were obtained through the Tissue Bank of Ghent University Hospital and Antwerp University Hospital under ethical approval of the Ethics Committee of Ghent University (2018/1072, B670201837286). Eye globes were provided with a description of time and cause of death, post-mortem circulation time, age and sex. None of the 6 donors had a prior known ophthalmological condition.  The eye globes were dissected on ice, followed by extraction of the neural retina and the retinal pigment epithelium/choroid. The resulting tissues were resuspended in 1XPBS supplemented with 10% Fetal Bovine Serum. The samples were processed according to Matelot & Noordermeer (2016) and cross-linking of nuclei was performed using 2% formaldehyde. Finally, the obtained nucleai were aliquoted per 10 million and snap frozen after supernatant removal. Samples were stored at -80°C. The generation of the 3C template was performed according to Schwartzman et al. 2016. Briefly, around 5 million cross-linked nuclei were digested overnight using 400 U DpnII (NEB). After digestion, ligation was performed overnight using 4,000 U of T4 DNA ligase (NEB), followed by the addition of proteinase K (BIOzymTC). Efficiency of digestion and ligation were evaluated via agarose gel electrophoresis. Next, samples were de-crosslinked, followed by purification of samples using AMPure XP beads (Agencourt). Subsequently, 4 µ