GSM8162176: nsn_input; Mus musculus; OTHER
Briefly, 200 oocytes freed of zona pellucida were incubated with concanavalin-coated magnetic beads (Vazyme, TD904) for 15 minutes at room temperature. CUT&Tag for H3K4me3 and H3K27me3 were performed following the protocol as previously described[45], with minor adaptations. Briefly, 200 oocytes freed of zona pellucida were incubated with concanavalin-coated magnetic beads (Vazyme, TD904) for 15 minutes at room temperature. This was followed by incubation with 1 μg of either H3K4me3 (Diagenode, C15410003) or H3K27me3 (Diagenode, C15410195) antibody at 4°C overnight, and subsequently with goat anti-rabbit IgG antibody (Vazyme, Ab207) for 1 hour. The samples were incubated with 0.08 μM pA/G-Tnp Pro on a rotator at room temperature for 1 hour, subsequently resuspended in 50 µL of Trueprep Tagment Buffer L, and incubated at 37°C for 1 hour. After tagmentation, DNA extraction was performed using DNA Extract Beads Pro. Finally, the libraries were constructed through 17 cycles of PCR amplification using the TruePrep Index Kit for Illumina (Vazyme, TD202).