Target nucleic acid sequence amplification method

C - Chemistry – Metallurgy – 12 – Q

Patent

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Details

C12Q 1/68 (2006.01) C07H 19/06 (2006.01) C07H 21/00 (2006.01) C12N 15/10 (2006.01) C12P 19/30 (2006.01) C12P 19/34 (2006.01)

Patent

CA 2262019

A target nucleic acid sequence amplification method wherein at least the target nucleic acid sequence, at least one oligonucleotide primer specific for the target sequence, one or more enzymatic activities and nucleotides are provided, and the target sequence is amplified under conditions suitable in particular for said enzymatic or activity or activities, is disclosed. According to the method, at least one of the nucleotides is a prefunctionalised nucleotide that differs from the other nucleotides at least in that it includes at least one unprotected reactive covalence function in at least one predetermined site of the base of said nucleotide, in order to give a prefunctionalised amplification product including at least one such prefunctionalised nucleotide. Nucleotides for carrying out the method are also disclosed.

L'invention concerne un procédé d'amplification d'une séquence d'un acide nucléique cible, selon lequel: on dispose au moins: de la séquence d'un acide nucléique cible, d'au moins une amorce oligonucléotidique spécifique de la séquence cible, d'une ou plusieurs activités enzymatiques, de nucléotides; on amplifie, dans des conditions adaptées notamment à l'activité ou les activités enzymatiques, la séquence cible, selon lequel au moins un des nucléotides est un nucléotide préfonctionnalisé, différent des autres nucléotides au moins par la présence d'au moins une fonction réactive de covalence, non protégée, disposée en au moins un site prédéterminé de la base dudit nucléotide, pour obtenir un produit d'amplification préfonctionnalisé comprenant au moins undit nucléotide préfonctionnalisé. L'invention concerne aussi des nucléotides pour mettre en oeuvre ce procédé.

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