GSM8852321: CutTag_HDAC1_V5 control_mitosis_rep1; Drosophila melanogaster; OTHER
Larval brains were dissected and dissociated into single cells, then the cells were arrested by ConA beads, and the proteins of interest were combined by the specific primary antibody, followed by secondary antibody and Tn5 transposase. To obtain interphase and mitosis cells, the dissociated cells were fixed with 0.5% formaldehyde for 8min and quenched by Glycine., then stained with pH3 (phospho-histone H3) and DAPI before ConA beads incubation. CUT&Tag libraries were prepared using Hyperactive Universal CUT&Tag Assay Kit for Illumina (Vazyme) complemented with TruePrep Index Kit V2 for Illumina (Vazyme). 2w-100w cells were used as starting materials. Libraries were amplified using 10-20 cycles on the thermocycler. Post-amplification libraries were purified using VAHTS DNA Clean Beads (Vazyme).