Hybridization assay using self-quenching fluorescence probe

  • US 6,030,787 A
  • Filed: 12/07/1998
  • Issued: 02/29/2000
  • Est. Priority Date: 11/16/1994
  • Status: Expired due to Term
First Claim
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1. A method for detecting a target polynucleotide in a sample comprising:

  • contacting said sample of nucleic acids with an oligonucleotide probe under conditions where said oligonucleotide probe selectively hybridizes to said target polynucleotide, said oligonucleotide probe including a fluorescent reporter molecule and a quencher molecule capable of quenching the fluorescence of said reporter molecule which are attached to said oligonucleotide probe such that said oligonucleotide probe is capable of adopting at least one single-stranded conformation when not hybridized to said target polynucleotide where said quencher molecule is fluorescent and quenches the fluorescence of said reporter molecule and is capable of adopting at least one conformation when hybridized to said target polynucleotide, where the fluorescence of said reporter molecule is unquenched such that the fluorescence intensity of said reporter molecule is greater than the fluorescence intensity of said quencher molecule when said oligonucleotide probe is hybridized to said target polynucleotide and said oligonucleotide probe is not hybridized with itself in the form of a hairpin structure; and

    monitoring the fluorescence of said reporter molecule under conditions where said oligonucleotide probe does not hybridize with itself to form a hairpin structure in order to detect the hybridization of said target polynucleotide to said oligonucleotide probe.

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