GSM3543873: mm_Late2cell_H3K4me3_rep1; Mus musculus; OTHER
CUT&RUN was conducted following the published protocol (Skene et al., 2018) with modifications.Oocytes or embryos removed zona pellucida were transferred into 0.2mL conventional, non-low-binding PCR tube (Axygen). Cells were attached to a solid support using concanavalin A–coated magnetic beads to allow simple handling. Following in situ binding of the antibody and pA-MN specifically to the target protein, pA-MN was activated by calcium at 0 °C. Cleaved fragments released with two cuts were free to diffuse out of the nuclei. After removing the intact nuclei by centrifugation, the supernatant containing released chromatin fragments were used to extract DNA directly for library construction. Purified DNA was subjected to Tru-seq library construction using either KAPA Hyper Prep Kit (KAPA biosystems, KK8502) or NEBNext Ultra II DNA Library Prep Kit for Illumina (NEB, E7645S).
GEO Accession
GSM3543873