Method for the high-parallel analysis of polymorphisms

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01) G01N 33/50 (2006.01)

Patent

CA 2411400

Disclosed is a method for the high-parallel characterisation of polymorphisms, especially SNPs. Said method can also be used for the simultaneous or separate detection of DNA methylation. First, a set of probes provided with at least one characteristic detectable identifying mark for a respective probe is connected to an addressed surface. A nucleic acid to be examined is then hybridised to these probes and the probes are extended in an allele-specific enzymatic reaction. The type and the occurrence of said allele-specific reaction determine whether the respective probe is enzymatically decomposed. Finally, the remaining allele-specific products are analysed and the existing alleles in the extracted nucleic acid samples are determined.

L'invention concerne un procédé servant à la caractérisation ultraparallèle de polymorphismes, notamment de SNP, et pouvant également être utilisé pour la détection simultanée ou distincte de la méthylation de l'ADN. Selon l'invention, on relie d'abord un jeu de sondes, pourvu d'au moins un repère détectable caractéristique de la sonde concernée, à une surface adressée. On hybride ensuite un acide nucléique à examiner avec ces sondes et on allonge les sondes lors d'une réaction enzymatique spécifique de l'allèle. Le type et l'existence de la réaction spécifique de l'allèle déterminent si la sonde concernée sera décomposée enzymatiquement lors de l'étape suivante. Enfin, on analyse les produits spécifiques de l'allèle restants et on procède à une détermination des allèles présents dans l'échantillon d'acide nucléique examiné.

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