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Quantitative detection of macromolecules with fluorescent oligonucleotides

  • US 5,445,935 A
  • Filed: 11/23/1992
  • Issued: 08/29/1995
  • Est. Priority Date: 11/23/1992
  • Status: Expired due to Fees
First Claim
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1. A method for measuring the binding of a polynucleotide to a macromolecule to form a complex which comprises:

  • i) labelling said polynucleotide with a fluorescent label;

    ii) obtaining a first measurement of the polarization of the fluorescent emission from said label;

    iii) contacting said labelled polynucleotide with said macromolecule, under conditions such that the concentration of said labelled polynucleotide ranges from 10 picomolar to 1 nanomolar;

    iv) obtaining a second measurement of the polarization of the fluorescent emission from said label;

    v) comparing said first measurement with said second measurement; and

    vi) detecting formation of said complex by observing an increase in the polarization measured in step (iv) compared to the polarization measured in step (ii);

    wherein said measurements of fluorescence polarization are made using an apparatus having a component selected from the group consisting of an apparatus having a Xenon arc lamp light source and a UV grade film polarizer, an apparatus having a Xenon arc lamp light source coupled by a fiber optic cable to a sample compartment and a UV grade film polarizer mounted directly on the sample compartment, and an apparatus having a laser light source.

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