GSM8187177: mESC, H3K9me3, UHRF1KO, rep1; Mus musculus; ChIP-Seq
Cells were fixed with 1% formaldehyde for 10 minutes and quenched by the addition of glycine to a final concentration of 125 mM for 5 min. Hypotonic lysis buffer and dounce homogenization was used to lyse cells. Nuclei were pelleted by centrifugation and sonicated to shear chromatin using Covaris E220 Focused-Ultrasonicator (140 watt peak incident, 5% duty factor, 200 cycles/burst, 330 secs sonication time) to a fragment distribution of 200 – 500 bp. Chromatin samples were incubated with specific antibodies overnight at 4°C. For H3K9me3 ChIP, 3 μg of anti-H3K9me3 (Abcam 8898) was used. For ChIP-seq, reverse crosslinked DNA was treated with RNase A and proteinase K. DNA was purified using phenol/chloroform/isoamyl alcohol and ethanol precipitated with 20 μg glycogen carrier. 10 ng of DNA was used to prepare libraries from two biological replicates per sample.