Retention of the native epigenome in purified mammalian chromatin
More about Open Access at the CrickAuthors list
Andreas H Ehrensberger Don-Marc Franchini Phil East Roger George Nik Matthews Sarah L Maslen Jesper SvejstrupAbstract
A protocol is presented for the isolation of native mammalian chromatin as fibers of 25-250 nucleosomes under conditions that preserve the natural epigenetic signature. The material is composed almost exclusively of histones and DNA and conforms to the structure expected by electron microscopy. All sequences probed for were retained, indicating that the material is representative of the majority of the genome. DNA methylation marks and histone marks resembled the patterns observed in vivo. Importantly, nucleosome positions also remained largely unchanged, except on CpG islands, where nucleosomes were found to be unstable. The technical challenges of reconstituting biochemical reactions with native mammalian chromatin are discussed.
Journal details
Journal PLOS ONE
Volume 10
Issue number 8
Pages e0133246
Available online
Publication date
Full text links
Publisher website (DOI) 10.1371/journal.pone.0133246
Figshare View on figshare
Europe PubMed Central 26248330
Pubmed 26248330