Methods and/or Use of Oligonucleotide Conjugates for Assays and Flow Cytometry Detections

A flow cytometry method for assaying a target of a sample, comprising:

• i) providing to the sample:
• 1) a molecular probe, comprising a binding moiety conjugated to a first oligonucleotide sequence;
• 2) a detectable component, comprising a signal generating moiety conjugated to a second oligonucleotide sequence;
• and
• 3) a universal adapter, comprising an oligonucleotide sequence having a first sequence segment complementary to the first oligonucleotide sequence of the molecular probe and a second sequence segment complementary to the second oligonucleotide sequence of the detectable component;
• ii) binding the target in the sample with the binding moiety of the molecular probe;
• iii) hybridizing the first oligonucleotide sequence of the molecular probe to the first oligonucleotide sequence segment of the universal adapter;
• iv) hybridizing the second oligonucleotide sequence of the detectable component to the second oligonucleotide sequence segment of the universal adapter;
• and
• v) detecting a signal generated from the hybridized detectable component using flow cytometry;
• wherein the method is characterized by one or more of the following:
• a) the conjugation between the first oligonucleotide sequence and the binding moiety and conjugation between the second 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 strong binding affinity for the target.