EpiScope Nucleosome Preparation Kit
The EpiScope Nucleosome Preparation Kit is designed for the preparation of nucleosomes from cultured mammalian cells. DNA is extracted from the prepared nucleosomes and analyzed using real-time PCR or sequencing to map nucleosome positions.
The EpiScope Nucleosome Preparation Kit is designed for the preparation of nucleosomes from cultured mammalian cells. DNA is extracted from the prepared nucleosomes and analyzed using real-time PCR or sequencing to map nucleosome positions.
Nucleosomes are the constituent units of chromatin in eukaryotic nuclei, consisting of genomic DNA wrapped around a core histone complex (Clark 2010). Nucleosomes are not spaced at regular intervals on genomic DNA; instead, their positions are dependent on chromatin structure and transcriptional regulation (Dutta and Workman 2012). In DNA methylation analysis, an important area in epigenetics research, nucleosome positioning has become a subject of intense interest. In transcriptionally active genes, the promoter region is free of nucleosomes, permitting the binding of transcriptional initiation factors (TIFs). Conversely, DNA methylation within gene promoter regions leads to the formation of nucleosomes in those regions, preventing the binding of TIFs and suppressing transcriptional initiation.
Overview
- Enables detailed analysis of nucleosome positioning
- Prepared DNA consists primarily of mononucleosomes
More Information
Applications
- Preparation of nucleosomal DNA from 0.5–2 x 106 mammalian cultured cells
- Nucleosome positioning with regards to DNA methylation analysis
Components
| Package 1 | |
|---|---|
| Micrococcal Nuclease (20 units/μl) | 25 ml × 2 |
| Micrococcal Nuclease Buffer (10X) | 1 ml |
| RNase A (20 mg/ml) | 100 μl |
| 0.5 M EDTA | 100 μl |
| Proteinase K (20 mg/ml) | 200 μl |
| LINE-1 qPCR Primer (10 μM each)* | 50 μl |
| Package 2 | |
|---|---|
| Cytoplasmic Lysis Buffer | 50 μl |
| Protease Inhibitor Cocktail (100X) | 550 μl |
*This component is to be used as a reference for making corrections to the amounts of genomic DNA in relative quantification analyses by real-time PCR. It is a primer for human genomic DNA detection and cannot be used with other species.
Storage
Package 1: –20℃
Package 2: 4℃
References
Clark, D. J. Nucleosome positioning, nucleosome spacing and the nucleosome code. J. Biomol. Struct. Dyn. 27, 781–93 (2010).
Dutta, A. & Workman, J. L. Nucleosome Positioning: Multiple Mechanisms toward a Unifying Goal. Mol. Cell 48, 1–2 (2012).
Additional product information
Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.
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