Retention of the Native Epigenome in Purified Mammalian Chromatin.
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...
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2015-01-01
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| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0133246 |
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| author | Andreas H Ehrensberger Don-Marc Franchini Philip East Roger George Nik Matthews Sarah L Maslen Jesper Q Svejstrup |
| author_facet | Andreas H Ehrensberger Don-Marc Franchini Philip East Roger George Nik Matthews Sarah L Maslen Jesper Q Svejstrup |
| author_sort | Andreas H Ehrensberger |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-1fe78f08e85f4033a0d681f092a4dba2 |
| institution | DOAJ |
| issn | 1932-6203 |
| language | English |
| publishDate | 2015-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-1fe78f08e85f4033a0d681f092a4dba22025-08-20T03:10:09ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-01108e013324610.1371/journal.pone.0133246Retention of the Native Epigenome in Purified Mammalian Chromatin.Andreas H EhrensbergerDon-Marc FranchiniPhilip EastRoger GeorgeNik MatthewsSarah L MaslenJesper Q SvejstrupA 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.https://doi.org/10.1371/journal.pone.0133246 |
| spellingShingle | Andreas H Ehrensberger Don-Marc Franchini Philip East Roger George Nik Matthews Sarah L Maslen Jesper Q Svejstrup Retention of the Native Epigenome in Purified Mammalian Chromatin. PLoS ONE |
| title | Retention of the Native Epigenome in Purified Mammalian Chromatin. |
| title_full | Retention of the Native Epigenome in Purified Mammalian Chromatin. |
| title_fullStr | Retention of the Native Epigenome in Purified Mammalian Chromatin. |
| title_full_unstemmed | Retention of the Native Epigenome in Purified Mammalian Chromatin. |
| title_short | Retention of the Native Epigenome in Purified Mammalian Chromatin. |
| title_sort | retention of the native epigenome in purified mammalian chromatin |
| url | https://doi.org/10.1371/journal.pone.0133246 |
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