Multichromophore fluorescent probes using DNA intercalation complexes
First Claim
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1. In a method for performing a separation of a composition comprising employing an electrical field for directing movement of components in said composition in a differential manner, the improvement which comprises:
- having present during said separation in said electrical field a moiety comprising (a) a probe having a double stranded DNA of at least about 20 base pairs and (b) non-covalently bound intercalated in said double stranded DNA at least one fluorescent molecule characterized by having at least two positive charges, having at least one fluorescent monomeric unit and having a binding affinity to double stranded DNA of at least about 5×
106 M−
1 and up to about 1 fluorescent unit per 4 nucleotides in a nucleic acid strand, wherein said fluorescent unit is characterized by having at least one aromatic ring per monomeric unit.
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Abstract
Novel fluorescent labeling techniques and fluorescent labels are provided, employing high affinity non-covalently binding and intercalating fluorescent dyes and dsDNA. The dyes find application to provide highly sensitive labeling of nucleic acids in electrophoretic gels and as pre-prepared labels for binding to a wide variety of specific binding pair members. The DNA-dye fluorescer complex can be used for labels in diagnostic assays, detection of specific nucleic acid sequences, and the like.
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Citations
10 Claims
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1. In a method for performing a separation of a composition comprising employing an electrical field for directing movement of components in said composition in a differential manner, the improvement which comprises:
having present during said separation in said electrical field a moiety comprising (a) a probe having a double stranded DNA of at least about 20 base pairs and (b) non-covalently bound intercalated in said double stranded DNA at least one fluorescent molecule characterized by having at least two positive charges, having at least one fluorescent monomeric unit and having a binding affinity to double stranded DNA of at least about 5×
106 M−
1 and up to about 1 fluorescent unit per 4 nucleotides in a nucleic acid strand, wherein said fluorescent unit is characterized by having at least one aromatic ring per monomeric unit.- View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9)
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10. The method according to claim 11, wherein said probe further comprises single stranded nucleic acid.
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