GSM2673314: A49 H3K9me3 ChIP-seq rep1; Mus musculus; ChIP-Seq
For ChIP-seq, ES cells were first fixed with 1% paraformaldehyde, and then lysed and sonicated to achieve the majority of DNA fragments at 100-1000 bp. DNA fragments were then enriched by immunoprecipitation with the H3K9me3 antibody (Abcam, ab8898). The immunoprecipitated material was eluted from the beads by heating for 30 min at 68 °C. Then, samples were incubated with Proteinase K at 42 °C for 2 h followed by 67 °C for 6 h to reverse the crosslinks. The samples were extracted with phenol:chloroform:isoamyl alcohol followed by chloroform, ethanol precipitation, and resuspension in TE buffer. For ChIP-seq, libraries were prepared according to Illumina protocols and library fragments of ~250 bp were used for sequencing.