Coupled amplification and ligation method

C - Chemistry – Metallurgy – 12 – Q

Patent

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C12Q 1/68 (2006.01)

Patent

CA 2195562

A method based on polymerase chain reaction (PCR) amplification and oligonucleotide ligase assay (OLA) reaction is provided for analyzing complex genetic systems in a single reaction vessel. The method involves simultaneously incubating a sample containing one or more target polynucleotides with PCR primers and OLA probes in a single reaction mixture. The presence of variant polynucleotide sequences in the sample is determined by detecting and identifying the products of the OLA reaction.

La présente invention concerne un procédé reposant sur une amplification par réaction en chaîne de la polymérase (PCR) et une réaction de dosage de ligases d'oligonucléotides (OLA). Ce procédé permet d'analyser des systèmes génétiques complexes dans une seule cuve à réaction. Le procédé consiste à faire incuber simultanément un échantillon contenant au moins un polynucléotide cible avec des amorces PCR et des sondes OLA dans un seul mélange de réaction. Pour déterminer la présence de séquences de polynucléotides variantes dans l'échantillon, le procédé consiste à détecter et identifier les produits issus de la réaction OLA.

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