GSM9312118: C&R, EyeDisc, IgG, Rep2; Drosophila melanogaster; OTHER
20 third-instar larval imaginal discs were dissected in Schneider medium at room temperature, centrifuged for 3 min at 700g and washed twice with wash+ buffer before addition of Concanavalin A-coated beads. CUT&RUN assays were performed according to Kami Ahmad's protocol implemented for Drosophila tissues (https://dx.doi.org/10.17504/protocols.io.umfeu3n) DNA libraries were prepared using the NEBNext® Ultra™ II DNA Library Prep Kit for Illumina (NEB). Adaptors were diluted 1:10 in water for adaptor ligation (step 2). The size selection of the adaptor-ligated DNA in step 3A was omitted (we proceeded directly to step 3B). We performed 14 cycles of PCR with 10 seconds of annealing/extension for enrichment of short DNA fragments. Libraries were sequenced on a NovaSeq 6000 system with 150 bp PE reads