A357C10-MP46A_H3K27me3
ChIP was performed from 6 millions of cells originating from Patient Derived Tumour grafts; magnetic Dynabeads coupled to Protein A were used for the IP (Invitrogen). Sonication was performed to obtain fragment size of 150-300 bp. Libraries were prepared accordingly to manufactures (TruSeq ChIP sample Prep Kit, Illumina). Sequencing was performed on a Illumina Hi-Seq 2500. Single-end 100bp reads were mapped on the Human reference genome using the Bowtie2 software (v2.1.0, Langmead B. et al. 2012) allowing one mismatch in the seed (22bp) and reporting one location in case of multiple mapping hits. PCR duplicates were then removed using PicardTools (v1.65, http://picard.sourceforge.net). Data was prepared for visualisation using HOMER (PMID: 20513432), by normalising the total number of tags is to 10 million. Furthermore, Peak calling was performed using MACS2 (pmid: 18798982) and differential binding was performed using DiffBind (PMID: 22217937).
Institut Curie
A357C10-MP46A_H3K27me3