Bio-Rad's new EpiQ chromatin analysis kit is a real-time PCR assay for the rapid quantitative assessment of chromatin structure.
Complementing existing epigenetic assays such as DNA methylation and chromatin
immunoprecipitation, the EpiQ kit is the first commercial research tool that
helps scientists quantify the impact of epigenetic events on gene expression
regulation through chromatin state changes.
The EpiQ kit can provide quantitative information about chromatin
accessibility, which correlates very strongly with gene expression.
Until now, the only way to quantitatively probe the status of chromatin
structure was through the use of homebrew methods, first described in 2001
(Rao, et. al., J. Immunol.). This approach requires the isolation and
purification of nuclei from several millions cells prior to chromatin
digestion and takes more than two days to complete. With the EpiQ kit,
chromatin structure data can be obtained within six hours from as few as
50,000 cultured cells, without the need for nuclei isolation.
The chromatin state of a gene can be identified in situ based on its
sensitivity to the action of the nuclease in the EpiQ kit. In
heterochromatin, genomic DNA is inaccessible to nuclease digestion and
remains available for subsequent qPCR. Analysis of heterochromatin using the
EpiQ kit reveals a minimal quantification cycle (Cq) shift between digested
and undigested samples. In contrast, DNA in euchromatin is susceptible to
nuclease digestion and is unavailable for qPCR. Analysis of euchromatin
using the EpiQ kit shows a large Cq shift between digested and undigested
EpiQ kit components include buffers for cell permeabilization and in situ
chromatin digestion, optimized nuclease, materials for genomic DNA
purification, control assays (qPCR primers) for chromatin assessment of a
reference (epigenetically silenced) and control (constitutively expressed)
gene, and EpiQ Chromatin SYBR® Green Supermix, a real-time PCR reagent
designed to amplify genomic DNA. The supermix can be used on all Bio-Rad
real-time PCR instruments and on those from other suppliers; some
instruments may require slight modifications to the supermix.
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