Single nucleotide amplification and detection by polymerase

Details Single nucleotide amplification and detection by polymerase Single nucleotide amplification and detection by polymerase

C - Chemistry, Metallurgy

12

Q

C12Q 1/68 (2006.01) C07H 19/00 (2006.01) C07H 19/10 (2006.01) C07H 19/20 (2006.01) C07H 21/00 (2006.01) C07H 21/02 (2006.01) C07H 21/04 (2006.01) C12P 19/34 (2006.01)

Patent

CA 2456906

A method of characterizing a nucleic acid sample is provided that includes the steps of: (a) conducting a DNA polymerase reaction that includes the reaction of a template, a non-hydrolyzable primer, at least one terminal phosphate- labeled nucleotide, DNA polymerase, and an enzyme having 3'.fwdarw.5' exonuclease activity which reaction results in the production of labeled polyphosphate; (b) permitting the labeled polyphosphate to react with a phosphatase to produce a detectable species characteristic of the sample; (c) detecting the detectable species; and (d) characterizing the nucleic acid sample based on the detection.

L'invention concerne une méthode de caractérisation d'un échantillon d'acide nucléique, consistant (a) à procéder à une réaction d'ADN polymérase comprenant la réaction d'un modèle, d'une amorce non hydrolysable, d'au moins un nucléotide marqué sur le phosphate de terminaison, d'ADN polymérase, et d'une enzyme à activité exonucléase 3'.fwdarw.5', ladite réaction générant la production d'un polyphosphate marqué; (b) à permettre au polyphosphate marqué de réagir avec une phosphatase afin d'obtenir une espèce détectable caractéristique de l'échantillon; (c) à détecter l'espèce détectable; et (d) à caractériser l'échantillon d'acide nucléique en fonction de cette détection.

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