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Methods and/or Use of Oligonucleotide Conjugates for Suppressing Background Due to Cross-Hybridization

  • US 20130344508A1
  • Filed: 05/20/2013
  • Published: 12/26/2013
  • Est. Priority Date: 04/18/2007
  • Status: Active Grant
First Claim
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1. A method for suppressing cross-hybridization in assaying one or more targets of a sample, comprising:

  • i) hybridizing;

    1) a molecular probe, comprising a binding moiety conjugated to an oligonucleotide sequence; and

    2) a detectable component, comprising a signal generating moiety conjugated to an oligonucleotide sequence complementary to the oligonucleotide sequence of the molecular probe;

    ii) providing to the hybridized molecular probe-detectable component a blocking component, comprising;

    1) an oligonucleotide sequence complementary to the oligonucleotide sequence of the molecular probe;

    or2) an oligonucleotide sequence complementary to the oligonucleotide sequence of the detectable component;

    iii) combining the blocked hybridized molecular probe-detectable component with one or more blocked hybridized molecular probe-detectable components similarly prepared by steps i) through ii);

    iv) combining the mixture of the two or more blocked hybridized molecular probe-detectable components and the sample comprising the one or more targets;

    v) binding the one or more targets in the sample with the binding moieties of the molecular probes of said two or more blocked hybridized molecular probe-detectable components;

    vi) detecting a signal generated from the one or more targets bound to the molecular probes of said two or more blocked hybridized molecular probe-detectable components;

    wherein the method is characterized by one or more of the following;

    a) the conjugation between the oligonucleotide sequence and the binding moiety and conjugation between the complementary oligonucleotide sequence and the signal generating moiety, comprises one or more covalent bond linkages, comprising a hydrazone, oxime, triazine, or other covalent bond, wherein the formation of the conjugates are at least 90% efficient; and

    b) the binding moiety comprises a binding affinity of less than 10

    4
    M for the target.

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