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Probes labeled with energy transfer couples dyes exemplified with DNA fragment analysis

  • US 5,869,255 A
  • Filed: 01/30/1997
  • Issued: 02/09/1999
  • Est. Priority Date: 02/01/1994
  • Status: Expired due to Term
First Claim
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1. A method of identification and detection of single or double stranded nucleic acids in a mixture of nucleic acids with different fluorescent labels covalently bonded to said nucleic acids to detect at least two different nucleic acids of interest having different mobilities in a separation means, wherein said labels are characterized by:

  • (1) having a molecular weight not exceeding about 10,000 daltons and comprising a donor-acceptor fluorescent pair bonded to a backbone chain with energy transfer from said donor to said acceptor; and

    (2) each of the labels absorbs at substantially the same wavelength and emits at a different wavelength, with the proviso that one of the labels may have the same fluorescer as the donor and acceptor;

    said method comprising;

    preparing a mixture of said nucleic acids wherein a portion of said nucleic acids are sample nucleic acids to which are bonded labels which emit light at one or more first wavelengths and a portion of said nucleic acids are standard molecular weight nucleic acids which have labels which emit light at one or more second wavelengths, said first and second wavelengths being different;

    separating or analyzing said mixture by said separation means as a result of different mobilities of said sample nucleic acids and said standard nucleic acids; and

    detecting each of said labeled nucleic acids by irradiating with a common light source at the absorption wavelength of said donor and detecting the fluorescence of each of said labels.

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