Patent ID: 6140054
Filing Date: 2000-10-31
Classification: C12Q

Abstract:
A method of analyzing a biological sample comprising a nucleic acid sequence for the presence of mutations or polymorphisms at multiple loci of the nucleic acid sequence, said method being conducted in a single reaction vessel and comprising the steps of(a) combining said biological sample with a pair of oligonucleotide PCR primers, a first donor oligonucleotide probe, a first acceptor oligonucleotide probe, a second donor oligonucleotide probe and second acceptor oligonucleotide probe,wherein said pair of oligonucleotide PCR primers is configured for amplifying a selected segment of the nucleic acid sequence; andsaid first and second donor oligonucleotide probes and said first and second acceptor oligonucleotide probes hybridize to the selected segment so that hybridization of both the first donor oligonucleotide probe and the first acceptor oligonucleotide probe to the selected segment places the first donor oligonucleotide probe and the first acceptor oligonucleotide probe in a resonance energy transfer relationship, and hybridization of both the second donor oligonucleotide probe and the second acceptor oligonucleotide probe to the selected segment places the second donor oligonucleotide probe and the second acceptor oligonucleotide probe in a resonance energy transfer relationship;(b) adding a thermostable polymerase and amplifying the selected segment of the nucleic acid sequence by the polymerase chain reaction;(c) illuminating the biological sample and monitoring the fluorescence as a function of temperature.