Self-quenching fluorescence probe and method

摘要

There is provided: a method for detecting a target polynucleotide in a sample comprising: contacting said sample of nucleic acids with an oligonucleotide probe under conditions favorable for hybridization; and monitoring the fluorescence of said reporter molecule, an increase in the fluorescence intensity of said reporter molecule indicating the presence of said target polynucleotide; wherein said oligonucleotide probe is an oligonucleotide probe comprising: an oligonucleotide probe which does not hybridize with itself to form a hairpin structure; a fluorescent reporter molecule attached to said oligonucleotide sequence; and a quencher molecule capable of quenching the fluorescence of said reporter molecule attached to said oligonucleotide sequence; said oligonucleotide sequence capable of existing in at least one single-stranded conformation when unhybridized where said quencher molecule is a fluorescent molecule capable of quenching the fluorescence of said reporter molecule; said oligonucleotide sequence also capable of existing in at least one conformation when hybridized to a target polynucleotide where the fluorescence of said reporter molecule is unquenched; and wherein the fluorescence intensity of said reporter molecule at its emission maximum when said oligonucleotide sequence is hybridized to said target polynucleotide is greater than the fluorescence intensity of said quencher molecule at its emission maximum when said oligonucleotide sequence is not hybridized to said target polynucleotide. <IMAGE>