GSM1348509: PFA-1; Homo sapiens; ChIP-Seq
10-20mg of fresh frozen primary tumour samples were homogenized in 1% Formaldehyde and allowed to incubate at room temperature for 6 – 10 min. Cross-linking was stopped with the addition of 125mM of Glycine, and samples were washed twice with ice-cold PBS containing (1% BSA and 10% FBS). Samples were then sonicated to ~200 bp fragments using a Biorupter (Diagenode). The chromatin immunoprecipitation was then performed using 5 mg of H3K27me3 antibody (C15410069-Diagenode) overnight at 4C as described previously (Lupien et al., Cell 2008). Following chromatin immunoprecipitation, DNA was quantified using picogreen, and libraries were prepared and barcoded according to NEB NEXT ChIP seq library preparation protocol performed using Agencourt bead purification. This protocol was followed according to NEB instructions.
GEO Accession
GSM1348509