A quantitative hybridization assay for the analysis of...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01)

Patent

CA 2447463

The invention provides a quantitative hybridization assay for the analysis of mRNA in a target nucleic acid sample. The method comprising the steps of (i) immobilizing the target nucleic acid sample on a solid support; (ii) contacting a labeled antisense probe to a first portion of the said target nucleic acid sample, and a labeled sense probe to a second portion of the said target nucleic acid sample; (iii) detecting and quantitating the signals generated from hybridized antisense probe and hybridized sense probe; and (iv) determining the value represented by the antisense probe signal minus the sense probe signal, said value being proportional to the amount of mRNA in the target nucleic acid sample.

L'invention a pour objet un dosage par hybridation quantitative en vue de l'analyse de l'ARNm dans un échantillon d'acide nucléique cible. Ce procédé comprend les étapes consistant à (i) immobiliser l'échantillon d'acide nucléique cible sur un support solide ; (ii) placer une sonde anti-sens marquée en contact avec une première partie de l'échantillon d'acide nucléique cible et une sonde sens marquée en contact avec une deuxième partie de l'échantillon d'acide nucléique cible ; (iii) détecter et quantifier les signaux générés à partir de la sonde anti-sens hybridée et la sonde sens hybridée et enfin (iv) déterminer la valeur représentée par le signal de sonde anti-sens moins le signal de sonde sens, cette valeur étant proportionnelle à la quantité d'ARNm dans l'échantillon d'acide nucléique cible.

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