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Abstract

An improved method for detecting, identifying and screening single polynucleotide polymorphisms, insertion/deletion loci, and microsatellites is provided. The method includes adding a donor intercalating dye to a sample containing an amplified target nucleic acid sequence, adding a probe containing an acceptor fluorophore to the sample, hybridizing the probe to the target sequence, exciting the donor dye with a specific wavelength of light, monitoring fluorescence from the sample due to FRET energy transfer from the dye to the probe fluorophore associated with one or both of the hybridization of the probe to the target sequence and the dissociation of the probe from the target sequence, and analyzing the sample using a melt-curve analysis to identify at least one single (or multiple) known or unknown nucleotide polymorphism, insertion/deletion loci, or microsatellite therein.

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First Claim