Method for nucleic acid analysis using fluorescence...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01)

Patent

CA 2240667

A method for detecting the presence of a target nucleotide or sequence of nucleotides in a nucleic acid is disclosed. The method is comprised of forming an oligonucleotide labeled with two fluorophores on the nucleic acid target site. The doubly labeled oligonucleotide is formed by addition of a singly labeled dideoxynucleoside triphosphate to a singly labeled polynucleotide or by ligation of two singly labeled polynucleotides. Detection of fluorescence resonance energy transfer upon denaturation indicates the presence of the target. Kits are also provided. The method is particularly applicable to genotyping.

L'invention concerne un procédé pour détecter la présence d'un nucléotide ou d'une séquence de nucléotides cibles dans un acide nucléique. Ce procédé consiste à former un oligonucléotide marqué par deux fluorophores sur le site cible d'acide nucléique. L'oligonucléotide à deux marqueurs est formé en ajoutant un didésoxynucléoside thriphosphate à un seul marqueur ou par ligation de deux polynucléotides à un seul marqueur. La détection du transfert des énergies de résonance par fluorescence lors de la dénaturation indique la présence de la cible. L'invention concerne également des kits. Le procédé s'applique particulièrement à la détermination des génotypes.

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