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Abstract

There is provided a method of amplifying a nucleic acid sequence. The method comprises providing an amplification mixture comprising an exonuclease capable of digestion of a strand of a double stranded nucleic acid molecule from the 5′-end towards the 3′-end, a strand displacing polymerase, a double stranded nucleic acid molecule comprising first and second nucleic acid strands, a first nucleic acid primer, and nucleotides as appropriate to provide for amplification of the first nucleic acid sequence to be amplified. The method further comprises effecting the amplification reaction under conditions permitting digestion, exonuclease digestion and strand displacement polymerisation thereby producing a product mixture comprising an amplified amount of said first nucleic acid sequence. There is also provided a method of determining the presence or quantity of target nucleic acid sequence in a biological sample using a double stranded probe having a fluorophore on one strand and its quencher on the other and using denaturation and re-hybridisation to detect the target.

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