GSM9207227: PS19-K9,rep3; Mus musculus; ChIP-Seq
Approximately 1× 10^7 cells from hippocampus as above was fixed in 1% formaldehyde for 10 min at room temperature (RT), followed by quenching in 125 nM glycine for 5 min. Cell nuclei were collected and lysed in lysing buffer (50 mM HEPES/KOH pH 7.6, 1 mM EDTA, 140 nM NaCl, 10% v/v Glycerol, 0.5% NP-40, 0.25% v/v Triton X-100) supplemented with protease inhibitor. Nuclei were subjected to sonication to acquire DNA fragments of 200-350 bp. The sonicated chromatin was applied to immunoprecipitation by incubation with 1.5 μg H3K9me3 antibody (Abcam, #ab232324) overnight at 4°C, followed with collection using 30 μl protein A/G plus agarose beads (Millipore). After elute and purify the chromatin, the ChIP-seq library was constructed using a KAPA HyperPrep Kits (Roche, 07962347001) according to the manufacturer's instructions，and then run on the Illumina sequencer Hiseq-Xten PE150. The ChIP libraries were prepared using KAPA HyperPrep Kits (Roche, 7962347001)