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Fluorescent labels and their use in separations

  • US 5,654,419 A
  • Filed: 02/01/1994
  • Issued: 08/05/1997
  • Est. Priority Date: 02/01/1994
  • Status: Expired due to Term
First Claim
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1. A method of separating nucleic acid components of a nucleic acid multicomponent mixture, wherein each of the different nucleic acid components or combinations of components of interest are labeled with different labels comprising different fluorescer pairs, wherein said labels are characterized by:

  • (1) having a donor-acceptor fluorescent pair bonded to an oligonucleotide chain with efficient energy transfer from said donor to said acceptor;

    (2) each of the donor members of the labels absorbs at substantially the same wavelength as all of the other donor members and each of the acceptor members of the labels emits at a different wavelength compared to any of the other acceptor members; and

    each of the different labels has substantially the same mobility in said separating as a result of varying the spacer of said donor-acceptor pair along said oligonucleotide chain, with the proviso that one of said labels can have the same fluorescer as the donor and the acceptor;

    said method comprising;

    binding different labels to different nucleic acid components or combinations of components of said multi-component mixture;

    separating said nucleic acid components into individual fractions; and

    detecting each of said labeled nucleic acid components by irradiating with a common light source at an absorption wavelength of said donors and detecting the fluorescence of each of said labels.

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