Method of quantitating genomic dna

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01) C12Q 1/70 (2006.01)

Patent

CA 2159043

A method of quantitating genomic DNA in a sample includes the steps of -) -) amplifying the DNA contained in the sample by a nucleic acid amplification process by using primers that are complementary to repetitive genomic sequences, and -) detecting the amplified DNA obtained; to obtain an exact quantitative information, -) a given amount of at least one known nucleic acid is added prior to amplification in a manner known per se to said sample as an internal standard, the standard nucleic acid differing from the genomic DNA to be quantitated in at least one detectable characteristic, and -) the amount of amplified genomic DNA and the amount of amplified standard nucleic acid are detected, and, departing from the amount of standard nucleic acid obtained, the amount of genomic DNA originally present in said sample is determined.

Une méthode de dosage de l'ADN génomique dans un échantillon comprend les étapes consistant à -) -) amplifier l'ADN contenu dans l'échantillon par un processus d'amplification des acides nucléiques au moyen d'amorces qui sont complémentaires aux séquences répétitives génomiques, et -) -) détecter l'ADN amplifié obtenu; pour obtenir une information quantitative exacte, -) une quantité donnée d'au moins un acide nucléique connu est ajouté avant l'amplification, de manière connue en soi, audit échantillon comme étalon interne, l'acide nucléique standard étant différent de l'ADN génomique à quantifier dans au moins une caractéristique détectable, et -) la quantité de l'ADN génomique amplifié et la quantité d'acide nucléique standard amplifié sont détectées, et, à partir de la quantité d'acide nucléique standard obtenu, on détermine la quantité d'ADN génomique initialement présent dans ledit échantillon.

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