GSM9168717: Sample 12_Photoreceptor Nuclei, LongGMR-Gal4 > UAS-Gnmt, H3K9me3, Replicate 1; Drosophila melanogaster; OTHER
200 male flies in triplicate or quadruplicate were collected at 10 days post eclosion and at Zeitgeber time 12 (ZT 12). GFP-positive photoreceptor nuclei were immunoprecipitated using the nuclei immunoenrichment protocol (dx.doi.org/10.17504/protocols.io.buiqnudw), followed by CUT&RUN (dx.doi.org/10.17504/protocols.io.zcpf2vn) with indicated antibodies (IgG - EpiCypher CUTANA IgG Negative Control Antibody, H3K9me3 - abcam # ab176916 (1 ug) , H3K27me3 - EpiCypher # 13-0055 (1 ug), H3K4me - abcam # ab8895 (.5 ug) ,H3K4me3 -EpiCypher # 13-0060 (1 ug)). IgG control samples use equivalent amounts of antibody with respective experimental samples. Samples were digested with MNase for 20 minutes. (0.4 ng - samples 1-19, or .65ng - samples 20-51) of DNA from each CUT&RUN reaction was used to prepare libraries using the NEBNext® Ultra™ II DNA Library Prep Kit for Illumina® from New England Biolabs.  Modifications to the library preparation were made to preserve short DNA fragments (dx.doi.org/10.17504/protocols.io.bagaibse)