title: GSM4827035: Monkey_16cell_PG_H3K27me3; Macaca fascicularis; ChIP-Seq
library_construction_protocol: H3K27me3 profling in early embryos are majorly based on a previously reported method cleavage under targests and release using nuclease (CUT&RUN). Briefly, several embryos containing total 200 ~ 300 cells were combined and transferred in a 1.5-ml tube containing 50 ul wash buffer. The embryos were then captured with BioMagPlus Concanavalin A, and incubated with a rabbit anti-H3K27me3 antibody for 16 hours at 4°C. After unbound antibodies were washed away, protein A-MNase was added and incubated for 3 hours at 4°C. After washes, CaCl2 was added to a final concentration of 2 mM to activate pA-MN, and the reaction was carried out for 20 min at 4°C and then stopped by addition of STOP buffer. The protein-DNA complexes were released by 10 min incubation at 37°C followed by centrifugation for 5 min at 16,000 g at 4°C. After transferring the supernatant to a new Lo-bound tube, SDS and Proteinase K were added and incubated at 55°C for at least 1 hour. DNA was then precipitated by phenol/chloroform/isoamylalcohol followed by ethanol precipitation with glycogen, and then dissolved in water. The purified DNA was prepared into library for sequencing using NEBNext UltraTM II DNA Libarary Prep Kit for Illumina according to manufacturer's instructions.