Methods and/or use of oligonucleotide conjugates for suppressing background due to cross-hybridization

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;
• and
• 2) a detectable component, comprising a signal generating moiety conjugated to an oligonucleotide having a sequence complementary to the sequence of the oligonucleotide of the molecular probe;
• ii) blocking the hybridized molecular probe-detectable component with a blocking component comprising
• 1) an oligonucleotide having a sequence complementary to the sequence of the oligonucleotide of the molecular probe, or
• 2) an oligonucleotide having a sequence complementary to the sequence of the oligonucleotide 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) to yield a mixture of the blocked hybridized molecular probe-detectable components;
• iv) combining the mixture of the blocked hybridized molecular probe-detectable components with the sample comprising the one or more targets;
• v) binding the one or more targets in the sample with one or more of the binding moieties of the molecular probes comprising the mixture of the blocked hybridized molecular probe-detectable components;
• vi) detecting a signal generated from the one or more targets bound to the one or more of the molecular probes that comprise the mixture of the blocked hybridized molecular probe-detectable components;
• wherein the method further is characterized by one or more of
• a) the conjugation between the oligonucleotide and the binding moiety and the conjugation between the complementary oligonucleotide and the signal generating moiety, wherein each conjugation comprises one or more covalent bond linkages comprising a hydrazone, oxime, triazine, or other covalent bond, wherein the formation of the conjugates is at least 90% efficient;
• and
• b) the binding moiety comprises a binding affinity for the target of less than 10−4 M.