GSM8240585: cSAM-control sg-H3K9me3 ChIP; Mus musculus; ChIP-Seq
All samples were carried out using SimpleChIP® Enzymatic Chromatin IP Kit (CST, 9003) following the manufacturer's instructions. 10^7 cultured cSAM cells per sample were fixed with 1% formaldehyde and then quenched with glycine. After washing and cell lysis, the chromatin was fragmented with 1 µl of micrococcal nuclease (MNase) at 37oC for 20 min and nuclei were completely lysed by sonication. The 10 µg of chromatin in each reaction was incubated with 10 µl of anti-H3K9me3 monoclonal antibody (CST, #13969) overnight at 4oC with rotation.  50 ng of spike-in chromatin (active motif, 53083) and 2 μg of spike-in antibody (active motif, 61686) were added to each ChIP reaction. Immunoprecipitation was performed with protein G magnetic beads. Following elution, reverse-crosslinks, and purification, DNA was used for further sequencing. The DNA fragments were repaired, A-tailed and further ligated with Illumina adapter. The final DNA library was obtained by size selection and PCR amplification.