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DETECTION AND QUANTIFICATION OF BIOMOLECULES USING MASS SPECTROMETRY

  • US 20080305479A1
  • Filed: 12/04/2007
  • Published: 12/11/2008
  • Est. Priority Date: 12/05/2006
  • Status: Active Grant
First Claim
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1. A method of detecting a target nucleic acid sequence in a sample, comprising the steps of:

  • (a) contacting a sample comprising a target nucleic acid with an (i) oligonucleotide primer, comprising a 3′

    end and a 5′

    end, and containing a sequence complementary to a region of the target nucleic acid and a (ii) detector oligonucleotide, comprising a 3′

    end and a 5′

    end, and containing a sequence complementary to a second region of the target nucleic acid sequence, thereby forming a (iii) mixture of duplexes under hybridization conditions, wherein the duplexes comprise the target nucleic acid annealed to the oligonucleotide primer and to the detector oligonucleotide such that the 3′

    end of the oligonucleotide primer is upstream of the 5′

    end of the detector oligonucleotide;

    (b) exposing the sample of step (a) to a cleavage agent under conditions sufficient to cleave and release the annealed detector oligonucleotide or at least one fragment thereof, thereby producing one or more mass-distinguishable products; and

    (c) detecting the one or more mass-distinguishable products by mass spectrometry, thereby detecting the presence or absence of the target nucleic acid sequence in the sample.

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