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Nucleic acid sequence detection employing probes comprising non-nucleosidic coumarin derivatives as polynucleotide-crosslinking agents

  • US 6,495,676 B1
  • Filed: 09/03/1999
  • Issued: 12/17/2002
  • Est. Priority Date: 04/13/1993
  • Status: Expired due to Fees
First Claim
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1. A method of detecting a target nucleic acid in a sample, said method comprising the steps of:

  • a) hybridizing a target nucleic acid to a crosslinkable probe, said crosslinkable probe comprising;

    i) a polynucleotide having a sequence complementary to that of said target nucleic acid;

    ii) a crosslinking moiety comprising a non-nucleosidic coumarin derivative incorporated into the backbone of said polynucleotide such that it replaces one or more nucleotides otherwise complementary to said target nucleic acid, wherein said coumain derivative is linked to a (poly)hydroxy hydrocarbon backbone moiety other than ribose or deoxyribose; and

    iii) optionally at least one label;

    b) activating said crosslinking moiety, whereby a covalent crosslink occurs between said probe and said target nucleic acid; and

    c) detecting the presence of a crosslinked nucleic acid pair as indicative of the presence of said target sequence in said sample.

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