Method for relative quantization of methylation of...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01)

Patent

CA 2378569

The invention concerns a method for relative quantization of methylation of cytosin-type bases in DNA samples, comprising the following steps: a) chemically reacting a genomic DNA sample with a reagent, 5-methylcytosin and cytosin reacting differently, thereby showing, after the reaction, different base-pairing behaviour in the DNA double helix; b) amplifying the DNA sample, a process during which a fluorescence-marked dCTP derivative is added; c) hybridizing the amplified DNA sample on at least an immobilised oligomer, the immobilised oligomers being each complementary to at least a primer used in the amplification step; and d) quantitatively measuring the fluorescence of the hybridized amplification product.

L'invention concerne un procédé de quantification relative de la méthylation des bases du type cytosine dans des échantillons d'ADN, qui comprend les étapes suivantes: a) on fait réagir chimiquement un échantillon d'ADN génomique avec un réactif, la 5-méthylcytosine et la cytosine réagissant de façon différente et montrant ainsi, après la réaction, un comportement d'appariement de bases différent dans la double hélice d'ADN; b) on amplifie l'échantillon d'ADN, opération pendant laquelle on ajoute un dérivé de dCTP marqué par fluorescence; c) on hybride l'échantillon amplifié d'ADN sur au moins un oligomère immobilisé, les oligomères immobilisés étant chacun complémentaires d'au moins une amorce utilisée à l'étape d'amplification; et d) on mesure quantitativement la fluorescence du produit d'amplification hybridé.

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