Isothermal strand displacement amplification using primers...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01) C12N 15/11 (2006.01)

Patent

CA 2609218

The invention is directed to a method for isothermal DNA amplification comprising providing to the DNA to be amplified an amplification mix comprising a first primer at least partially complementary to a region of DNA and containing a non-regular base, a second primer at least partially complementary to a region of DNA and containing a non- regular base, a DNA polymerase, an enzyme capable of strand displacement, an enzyme that recognises a non-regular base in double-stranded DNA and causes a nick or excises a base in one DNA strand at or near the non-regular base; and amplifying the DNA substantially without thermal cycling.

L'invention concerne un procédé pour l'amplification d'ADN isotherme consistant à fournir à l'ADN à amplifier un mélange d'amplification comprenant une première amorce au moins partiellement complémentaire à une région d'ADN et contenant une base non régulière, une deuxième amorce au moins partiellement complémentaire à une région d'ADN et contenant une base non régulière, un ADN polymérase, une enzyme capable de déplacement de brin, une enzyme qui reconnaît une base non régulière dans un ADN à double brin et provoque une coupure simple brin ou excise une base dans un brin d'ADN au niveau de ou près de la base non régulière ; et l'amplification de l'ADN sensiblement sans cycle thermique.

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