Methods for quantitating small rna molecules

C - Chemistry – Metallurgy – 12 – Q

Patent

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C12Q 1/68 (2006.01)

Patent

CA 2693974

In one aspect, the present in-vention provides methods, compositions and kits for amplifying a microRNA molecule to produce DNA molecules. The methods each include the steps of: (a) using primer exten-sion to make a DNA molecule that is comple-mentary to a target microRNA molecule; and (b) using a universal forward primer and a re-verse primer to amplify the DNA molecule to produce amplified DNA molecules. In some embodiments of the method, at least one of the forward primer and the reverse primer com-prise at least one locked nucleic acid molecule.

La présente invention propose, sous un aspect, des procédés, des compositions et des kits pour amplifier une molécule de microARN pour produire des molécules d'ADN. Les procédés consistent chacun: (a) à utiliser une extension d'amorce pour produire une molécule d'ADN qui est complémentaire d'une molécule de microARN cible; et (b) à utiliser une amorce avant universelle et une amorce inverse pour amplifier la molécule d'ADN pour produire des molécules d'ADN amplifiées. Dans certains modes de réalisation du procédé, l'amorce avant et/ou l'amorce inverse comporte au moins une molécule d'acide nucléique bloquée.

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