Method for identifying nucleic acid molecules

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01) C12N 15/10 (2006.01)

Patent

CA 2328146

A method of analyzing a DNA molecule is disclosed. In one embodiment the method comprises the steps of exposing a DNA molecule to an effective amount of a chemical modification reagent wherein the reagent converts guanine to 8- hydroxyguanine. The oxidized product is then exposed to a DNA glycosylase enzyme and the DNA molecule is cleaved at the site of the 8-hydroxyguanine. The fragments are then resolved by electrophoresis and the position of guanine residues within the DNA molecule is determined. In a preferred embodiment of the present invention, the modification reagent is a thiazine dye and the enzyme is FPG protein.

La présente invention concerne un procédé d'analyse de molécule d'ADN. Dans une réalisation, le procédé comprend plusieurs étapes dont la première consiste à exposer une molécule d'ADN à une quantité suffisante de réactif de modification chimique, ce réactif transformant la guanine en 8-hydroxyguanine. Le produit oxydé est alors exposé à une enzyme ADN-glycosylase qui coupe la molécule d'ADN à l'endroit contenant la 8-hydroxyguanine. Les fragments sont séparés par électrophorèse, ce qui permet de déterminer la position des résidus guanine dans la molécule d'ADN. Dans une réalisation préférée, le réactif de modification est un colorant thiazine et l'enzyme est une protéine FPG.

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