Method for amplifying specific nucleic acid sequences

C - Chemistry – Metallurgy – 12 – Q

Patent

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C12Q 1/68 (2006.01)

Patent

CA 2218026

The present invention provides a method of detecting a genetic polymorphism in an individual. In one form the method comprising the following steps: (1) obtaining a sample containing nucleic acid from the individual; (2) amplifying the nucleic acid sample from step (1) by a process involving thermocycling and primers, the amplification occurring in the presence of a thermostable restriction endonuclease which retains activity during thermocycling, the primers being selected such that they introduce into either the nucleic acid amplified from nucleic acid not including the polymorphism or from nucleic acid including the polymorphism, a sequence recognised by the thermostable restriction endonuclease; and (3) analyzing the product of step (2) to determine the presence or absence of the polymorphism.

La présente invention concerne un procédé pour détecter un polymorphisme génétique chez un individu. Sous une forme, ledit procédé consiste à: (1) obtenir un échantillon contenant de l'acide nucléique dudit individu; (2) amplifier l'échantillon d'acide nucléique obtenu à l'étape (1) au moyen d'un procédé faisant intervenir le thermocyclage et des amorces, l'amplification se faisant en présence d'une endonucléase de restriction thermostable qui conserve son activité au cours du thermocyclage, les amorces étant sélectionnées de façon à ce qu'elles introduisent une séquence reconnue par l'endonucléase de restriction thermostable soit dans l'acide nucléique amplifié à partir de l'acide nucléique ne comprenant pas le polymorphisme, soit dans l'acide nucléique amplifié à partir de l'acide nucléique comprenant le polymorphisme; et (3) analyser le produit résultant de l'étape (2) afin de déterminer la présence ou l'absence du polymorphisme.

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