Method of labeling a nucleic acid amplicon with simultaneous contamination prevention
AssigneeApplied Gene Technologies, Inc.
InventorsNanibhushan Dattagupta, C. Nagaraja Sridhar, Whei-Kuo Wu
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Abstract
This invention relates to methods of amplifying nucleic acids to minimize contamination by products of earlier amplification reactions. More particularly, it relates to methods of using nucleic acid labels that inhibit further amplification of the amplicon, and compositions that are useful to accomplish this task. In particular, the present invention relates to photoreactive complexes of a binding ligand, a binding enhancer and a label.
Claims45
A complex for labeling a nucleic acid in a sample comprising:
- a photoreactive binding ligand comprising a chemical moiety that binds to a nucleic acid and that, when activated by light, forms at least one covalent bond therewith;
- a binding enhancer comprising a chemical moiety that has a specific affinity for nucleic acids;
- and
- a label comprising a detectable chemical moiety;
- wherein said binding ligand, binding enhancer and label are bound together in the form of a covalent or noncovalent complex.
A composition for labeling and decontaminating a nucleic acid amplification reaction product comprising at least two intercalator moieties and a detectable label, wherein at least one of said intercalator moieties forms at least one covalent bond with said nucleic acid amplification reaction product.
A complex for labeling a nucleic acid in a sample comprising:
- a photoreactive binding ligand comprising a chemical moiety that binds to a nucleic acid and that, when activated by light, forms at least one covalent bond therewith;
- a binding enhancer comprising a chemical moiety that has a specific affinity for nucleic acids;
- and
- a label comprising a detectable moiety;
- and
- at least one lysis reagent;
- wherein said binding ligand, binding enhancer and label are bound together in the form of a covalent or noncovalent complex.
A method for labeling a nucleic acid in a clinical sample, comprising the steps of:
- adding a photoreactive binding ligand to the clinical sample, wherein the binding ligand comprises a chemical moiety that binds to a nucleic acid and, when activated by light, forms at least one covalent bond therewith;
- adding a binding enhancer to the clinical sample, wherein the binding enhancer comprises a chemical moiety that has a specific affinity for nucleic acids;
- and
- adding a label comprising a detectable moiety;
- and
- at least one lysis reagent;
- wherein said binding ligand, binding enhancer and label are bound together in the form of a covalent or noncovalent complex.
Dependencies are inferred from the claim text.