Differential amplification of mutant nucleic acids by pcr in...

C - Chemistry – Metallurgy – 12 – Q

Patent

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Details

IPC codes

C12Q 1/68 (2006.01)

Type

Patent

Patent number

CA 2635109

Description

A method for enriching a mutant nucleic acid in a mixture of nucleic acids, wherein the method comprises: (a) providing a nucleic acid mixture comprising a parental nucleic acid and a mutant nucleic acid of the parental nucleic acid; and (b) amplifying the nucleic acids in the nucleic acid mixture by polymerase chain reaction (PCR); wherein the mutant nucleic acid is a G.fwdarw.A mutant of the parental nucleic acid, which pairs with a fully complementary nucleic acid sequence to form an AT-rich nucleic acid variant of the parental nucleic acid; and wherein the AT-rich nucleic acid variant is denatured and selectively amplified by carrying out PCR using a denaturation temperature 1-3~C lower than the lowest denaturation temperature (Tp) that allows amplification of the parental nucleic acid to thereby enrich the mutant nucleic acid in the nucleic acid mixture.

L'invention concerne un procédé permettant d'enrichir un acide nucléique mutant dans un mélange d'acides nucléiques. Ce procédé consiste à: (a) prendre un mélange d'acide nucléique contenant un acide nucléique parental et un acide nucléique mutant de cet acide nucléique parental, et (b) amplifier les acides nucléiques présents dans le mélange d'acides nucléiques par réaction en chaîne de la polymérase (PCR). L'acide nucléique mutant est un mutant G.fwdarw.A de l'acide nucléique parental qui s'apparie à une séquence d'acide nucléique entièrement complémentaire pour former un variant d'acide nucléique riche en AT de l'acide nucléique parental, et ce variant d'acide nucléique est dénaturé et amplifié sélectivement par PCR à une température de dénaturation inférieure de 1 à 3 °C à la température de dénaturation la plus basse (Tp) permettant l'amplification de l'acide nucléique parental, afin d'enrichir l'acide nucléique mutant dans le mélange d'acide nucléique.

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