GSM8395537: znf706_CT_rep1; Danio rerio; OTHER
The operation from the anesthetization of adult fish to oocyte digestion is the same as that for RNA-seq. It is worth noting that CUT&Tag is a technique capturing genomic DNA, thus, it is imperative to eliminate the contamination from the granulosa cells and other somatic cells that envelop the oocytes. In our study, we performed the digestion by cell dissociation buffer (PCM, PC-33689) to get rid of somatic cells. The oocyte surface exhibits a pronounced smoothness, signifying the removal of somatic cells, which would be further validated by DAPI staining. Then, the medium containing the oocytes was drawn through a 100 ?m cell strainer using a pipette, followed by a 2-minute incubation period to allow for the gravitational sedimentation of the oocytes at the bottom of the plate. To remove impurities, the sample was thoroughly washed 2-3 times with a preheated L-15 medium. Approximately 5,000 WT stage I oocytes were collected, followed by library construction using the Hyperactive U  niversal CUT&Tag Assay Kit for Illumina Pro (Vazyme, TD904) and sequencing (Novogene, Hiseq-PE150 platform). The primary antibody used in the experimental group is rabbit anti-Znf706 (Aviva, arp32987-P050, 2.5?g / 50?l), while rabbit anti-IgG antibody (Cell Signaling Technology, 2729s, 2.5 ?g / 50?l) in the control group. The secondary antibody was goat anti-rabbit IgG H&L (Abcam, ab6702, 1:100 dilution). The experimental group underwent two biological replicates.