DNA detection by color complementation

  • US 5,489,507 A
  • Filed: 05/01/1991
  • Issued: 02/06/1996
  • Est. Priority Date: 11/30/1988
  • Status: Expired due to Term
First Claim
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1. A method of determining the zygosity of an individual at a predetermined genetic locus having one or more allelic forms of DNA, the method comprising the steps of:

  • providing a labelling means for each of the one or more allelic forms of DNA, such that each labelling means comprises a pair of oligonucleotides, each oligonucleotide of the pair being capable of forming perfectly matched duplexes at different and nonoverlapping regions of the same allelic form of DNA, and such that each labelling means is capable of producing a light signal different from every other labelling means;

    combining the labelling means with the sample;

    simultaneously amplifying the allelic forms of DNA forming perfectly matched duplexes with the pairs of oligonucleotides of their respective labelling means;

    separating from the amplified allelic forms of DNA, any labelling means having an oligonucleotide incapable of forming a perfectly matched duplex with any of the allelic forms of DNA; and

    illuminating the amplified allelic forms of DNA with an illumination beam such that a distinct light signal is produced by color complementation of the light signals generated by the labelling means associated with the amplified allelic forms of DNA.

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