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The EpiMark® Nucleosome Assembly Kit is used to make unmodified recombinant human nucleosomes with user-supplied DNA or the provided control DNA. Purified recombinant human Histone H2A/H2B Dimer and Histone H3.1/H4 Tetramer are mixed with DNA at 2 M NaCl. To generate nucleosomes, the salt concentration is lowered by dilution or dialysis allowing each histone tetramer to associate with two histone dimers and form the histone octamer on the DNA (1,2). A method for assaying nucleosome formation by gel shift assay is included. These nucleosomes may serve as better substrates for enzymes that are inactive on the DNA or one of the core histones alone (3,4). Each reaction creates nucleosomes from ~1 µg of 208 bp DNA and may be scaled depending on the experiment.
Nucleosome Assembly
A gel shift assay allows visualization of complex formation when various ratios of Octamer* to DNA are used to form mononucleosomes using the provided nucleosome control DNA. When assembly reactions are run on 6% polyacrylamide gel in 0.5X TBE, a characteristic shift occurs from 208 bp to approximately 700 bp.
M: Low Molecular Weight DNA Ladder (NEB #N3233)
Lane 1: Nucleosome Control DNA
Lane 2: 0.5:1 ratio of Octamer* to DNA
Lane 3: 1:1 ratio of Octamer* to DNA.
*Octamer = 2:1 mix of Histone H2A/H2B Dimer and Histone H3.1/H4 Tetramer.
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The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
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This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.
Trademarks
EPIMARK® is a registered trademark of New England Biolabs, Inc. TRIDYE™ is a trademark of New England Biolabs, Inc.
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