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Hybridization of polynucleotides conjugated with chromophores and fluorophores to generate donor-to-donor energy transfer system

  • US 6,162,603 A
  • Filed: 07/28/1998
  • Issued: 12/19/2000
  • Est. Priority Date: 11/07/1991
  • Status: Expired due to Fees
First Claim
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1. A method for detection of a target polynucleotide sequence, comprising the steps of:

  • providing a first polynucleotide sequence having one or more chromophores;

    providing a second polynucleotide sequence hybridized to the first polynucleotide sequence, the second polynucleotide sequence having one or more chromophores arranged in a quenching relationship to the acceptor chromophore of the first polynucleotide sequence when the first polynucleotide sequence and second polynucleotide sequence are hybridized;

    exposing a target polynucleotide sequence to the hybridized first and second polynucleotide sequences;

    denaturing the hybridized first and second polynucleotide sequences;

    hybridizing the first polynucleotide sequence to the target polynucleotide sequence;

    hybridizing a third polynucleotide sequence having one or more donor chromophores to the target polynucleotide sequence; and

    irradiating the mixture to detect hybridization of the first polynucleotide sequence to the target polynucleotide sequence by fluorescence energy transfer from the one or more donor chromophores of the third polynucleotide sequence to the one or more acceptor chromophores of the first polynucleotide sequence.

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