GSM8677809: FGO_control_H3K27me3_rep2; Mus musculus; ChIP-Seq
8-week-old female mice were intraperitoneally injected with PMSG. Oocytes were harvested in M2 medium 48 h after PMSG. STAR ChIP-seq was performed following the previously described protocol(Zhang et al., 2016). Briefly, samples were directly lysed and fragmented by MNase at 37°C for 5 minutes, before being incubated with the primary antibody (H2A.Z, abcam, 4174;H3K36me3, Active Motif, 61021; H3K4me3(Zhang et al., 2016); H3K27me3, Diagenode, C15410069; H3K27ac, Active Motif, 39133) overnight with rotation at 4°C. The next day, 100 μg protein A dynabeads (Life Technologies) were added to each sample and incubated for 2-3 h with rotation at 4 °C. The beads were washed five times with 150 μl RIPA buffer and once with 150 μl LiCl buffer. For each sample, beads were resuspended with 27 μl ddH2O and 1 μl 10× Ex-Taq buffer (TaKaRa). 1 μl proteinase K (Roche, 10910000) was added to each sample and incubated at 55 °C for 90 min to elute DNA from beads. After incubation at 80°C for 40min to inactivate the proteinase K, individual sample was subjected to TELP library preparation without DNA purification