Methods for quantitating small rna molecules

C - Chemistry – Metallurgy – 12 – Q

Patent

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C12Q 1/68 (2006.01)

Patent

CA 2595716

In one aspect, the present invention provides methods for amplifying a microRNA molecule to produce DNA molecules. The methods each include the steps of: (a) using primer extension to make a DNA molecule that is complementary to a target microRNA molecule; and (b) using a universal forward primer and a reverse 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 comprise at least one- locked nucleic acid molecule.

Dans un mode de réalisation, l'invention concerne des procédés pour amplifier une molécule de microARN pour produire des molécules d'ADN. Ces procédés comprennent respectivement les étapes qui consistent : (a) à utiliser une extension d'amorce pour générer une molécule d'ADN qui est complémentaire à une molécule de microARN cible, et ; (b) à utiliser une amorce sens universelle et une amorce antisens pour amplifier la molécule d'ADN pour produire des molécules d'ADN amplifiées. Dans certains modes de réalisation, au moins l'amorce sens ou l'amorce antisens comprend au moins une molécule d'acide nucléique verrouillé.

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