GSM4041187: CRISPR_mut_enhancer_rep1; Drosophila melanogaster; OTHER
in nucleus Hi-C libraries were generated for S2R+ wild-type cells and CRISPR mutants at least in duplicates using an in nucleus Hi-C protocol with minor modifications for S2R+ cells (Rubín et al 2017). Briefly, 40x106 S2R+ cells were fixed with formaldehyde at a final concentration of 2% for 10 minutes at room temperature and fixing reaction was stopped by adding 1M Glycine for 5 minutes rocking. Cells were washed with cold-PBS and flash frozen for short-term storage at -70°C or used immediately for subsequent experiments. The genomes were digested with MboI (NEB) and 5' overhangs were filled with Biotin-14-dATP (Invitrogen). Hi-C libraries were sequenced at the Babraham Institute sequencing facility.