Multiplex standardized reverse transcriptase-polymerase...

C - Chemistry – Metallurgy – 12 – P

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C12P 19/34 (2006.01) C12Q 1/00 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2480160

A method for direct comparison of numeral gene expression values between samples of genes using reverse transcription-polymerase chain reaction is described. CDNA, a competitive template mixture, and primer pairs for a plurality of genes are combined with at least one suitable buffer and at least one suitable enzyme to form a mixture. The mixture is amplified for a predetermined number of cycles to form PCR products. The PCR products are mixed with at least one suitable buffer, at least one enzyme, and one primer pair specific for each of the genes. The resulting mixture is amplified an additional predetermined number of cycles.

L'invention concerne un procédé de comparaison directe de valeurs numériques d'expression génique parmi des échantillons de gènes à l'aide de l'amplification en chaîne par polymérase de transcription inverse. On combine l'ADNc, un mélange de matrice compétitif, et des paires d'amorce pour une pluralité de gènes avec un tampon adéquat au moins et une enzyme adéquate au moins afin d'obtenir un mélange. On amplifie le mélange pendant un certain nombre de cycles afin d'obtenir des produits PCR. On mélange ensuite ces produits avec un tampon adéquat au moins, une enzyme au moins et une paire spécifique d'amorces pour chaque gène. On mélange enfin le mélange ainsi obtenu pendant un nombre prédéterminé supplémentaire de cycles.

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