Patent ID: 6395518
Filing Date: 2002-05-28
Classification: C07H,C12Q,Y10S

Abstract:
A method for performing combined PCR amplification and hybridization probing comprising the steps of:contacting a target nucleic acid sequence with PCR reagents, including at least two PCR primers and a polymerase enzyme substantially lacking any 5â€²â†’3â€² exonuclease activity, and an oligonucleotide probe comprising: an oligonucleotide; a fluorescer molecule attached to a first end of the oligonucleotide; a quencher molecule attached to a second end of the oligonucleotide such that quencher molecule substantially quenches the fluorescence of the fluorescer molecule wherein the oligonucleotide probe is in a single-stranded state and such that the fluorescer is substantially unquenched whenever the oligonucleotide probe is in a double-stranded state; and a 3â€² end which is rendered impervious to the 5â€²â†’3â€² extension activity of a polymerase; and subjecting the target nucleic sequence, the oligonucleotide probe, and the PCR reagents to thermal cycling sufficient to amplify the target nucleic acid sequence specified by the PCR reagents.