Accurate and efficient quantification of dna sensitivity by...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01) C07H 21/04 (2006.01) C12N 9/12 (2006.01) C12P 19/34 (2006.01)

Patent

CA 2448757

The sensitivity of regions of genomic DNA from chromatin to DNA-modifying agents are quantitated using real time PCR with careful use of reference regions and standard curves for improved accuracy over previous methods. A wide variety of DNA modifying agents are used to modify DNA, including single strand breakers, which allows analysis of a wider variety of DNA structures with greater precision than was previously possible. Methods also are provided that allow fine detailed analysis of hypersensitive DNA regions from as little as 100 cells. These methods also potentiate the investigation of more fine details of chromatin fiber folding and nucleosome-nucleosome contact.

L'invention concerne la quantification de la sensibilité de régions d'ADN génomique de chromatine à des agents de modification de l'ADN par utilisation de PCR en temps réel avec une utilisation prudente de régions de référence et de courbes témoin aux fins d'amélioration de la précision par rapport aux procédés antérieurs. On utilise une large variété d'agents de modification d'ADN afin de modifier l'ADN, y compris des agents de rupture de simple brin, permettant l'analyse d'une variété plus importante de structures d'ADN avec une meilleure précision qu'auparavant. L'invention concerne aussi des procédés permettant une analyse fine détaillée de régions d'ADN hypersensibles aussi petites que 100 cellules. Ces procédés favorisent aussi la recherche de détails plus fins du repliement de fibre de chromatine et de contact entre nucléosomes.

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