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- Nucleosome Mobilization Assay (Epigenome Network of Excellence)
The organization of eukaryotic genomes into nucleosome arrays restricts DNA sequence accessibility to many nuclear factors. Thus most DNA-based processes require opening (or "re-closing") of these arrays. One major class of enzymes, the "chromatin/nucleosome remodeling" factors, uses ATP hydrolysis to alter the canonical histone-DNA contacts. The term "nucleosome remodeling" can be defined and monitored in different ways. The simplest configuration to study one aspect of nucleosome remodeling is to use a purely reconstituted system consisting of mononucleosomes and an ATP-dependent nucleosome remodeler in the so-called "nucleosome sliding" or "nucleosome mobilization" assay. This technique was initially developed by Carl Wu and Peter Becker laboratories by taking advantage of two nucleosome properties: Nucleosomal histones can moderately move on DNA under rather mild temperature and salt conditions and nucleosomes reconstituted on a short DNA fragment can adopt multiple positions that can be separated by native gel electrophoresis
http://www.epigenome-noe.net/researchtools/protoco...
Added: Wed Feb 04 2009, Hits: 279, Reviews: 0 Write review Cached -
In vitro reconstitution of nucleosome arrays with a stoichiometric content of histone octamer and li
(Epigenome Network of Excellence)
To overcome the limitations of using native chromatin samples, we have developed an in vitro nucleosome array reconstitution system that produces very long, highly regular and soluble nucleosome arrays, or folded ‘30nm’ chromatin fibres, with a stoichiometry of one histone octamer and up to one linker histone per nucleosome.
http://www.epigenome-noe.net/researchtools/protoco...
Added: Wed May 27 2009, Hits: 187, Reviews: 0 Write review Cached -
Nucleosome Mapping using Mnase
(Tsukiyama Lab, Fred Hutchinson Cancer Research Center)
http://labs.fhcrc.org/tsukiyama/protocols/nucleoso...
Added: Tue Mar 03 2009, Hits: 1019, Reviews: 0 Write review -
Nucleosome Spacing Assay
(Tsukiyama Lab, Fred Hutchinson Cancer Research Center)
http://labs.fhcrc.org/tsukiyama/protocols/nucleoso...
Added: Tue Mar 03 2009, Hits: 803, Reviews: 0 Write review -
Site-Directed Hydroxy Radical Mapping of Nucleosome Positions in Vitro
(Epigenome Network of Excellence)
This protocol gives details of a method for mapping the position of nucleosomes in vitro by attaching an EDTA-derived reagent to a specific site on the histone octamer which then catalyses local DNA cleavage. The sites of cleavage reveal the position of the nucleosome at base pair resolution. Nucleosome positions and movements on DNA fragments of some 500 bp have been mapped
http://www.epigenome-noe.net/researchtools/protoco...
Added: Wed Feb 04 2009, Hits: 173, Reviews: 0 Write review Cached