GSM5531329: mouse_G9a_Ctrl_H3K9me2_rep2; Mus musculus; ChIP-Seq
Samples were collected in Nuclei Extraction Buffer directly and sheared by micrococcal nuclease (MNase) at 25°C for 5 min. Then the samples were the was incubated with H3K9me2 or CTCF antibody overnight with rotation under constant rotation on a rotator at 4°C. The next day, 10ul Dyna beads Protein A (Thermo Fisher Scientific) was added to each sample and incubated 2 hours up to overnight at 4 °C. Subsequently, the beads were washed twice with low salt wash buffer, twice high salt wash buffer. For each sample, 100 ul eluiton buffer were added to resuspend beads and incubated at 65℃ for 2h to elute DNA from the beads. The DNA was purified by phenol:chloroform:isoamyl alcohol method. Then samples were subjected to CUT&RUN  or STAR-seq library preparation. ULI-NChIP library was generated using the KAPA Hyper Prep Kit according to the manufacture's protocol. Paired-end 150-bp sequencing was performed on a HiSeq2500.