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Products >  Epigenetics >  DNA_Methylation_Analysis >  EpiScope_Nucleosome_Preparation_Kit

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.

Nucleosomes are the constituent units of chromatin in eukaryotic nuclei, consisting of genomic DNA wrapped around a core histone complex1. Nucleosomes are not spaced at regular intervals on genomic DNA; instead, their positions are dependent on chromatin structure and transcriptional regulation2. 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.

At-A-Glance Documents Images & Data

Features

  • Enables detailed analysis of nucleosome positioning
  • Prepared DNA consists of primarily mononucleosomes

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 (10×)1 ml
RNase A (20 mg/ml)100 μl
0.5 M EDTA100 μl
Proteinase K (20 mg/ml)200 μl
LINE-1 qPCR Primer (10 μM each)*50 μl

Package 2

Cytoplasmic Lysis Buffer50 μl
Protease Inhibitor Cocktail (100×)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

1. Clark, D.J. (2010) J. Biomol. Struct. Dynamics 27(5):781-793.

2. Dutta, A. & Workman, J.L. (2012) Mol. Cell 48(1):1-2.




 
 
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