Intercalation inhibition assay for compounds that interact with DNA or RNA
First Claim
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1. A process for the quantitative determination of the extent of binding of a compound to a nucleic acid, which comprises the steps:
- (a) preparing a mixture containing said compound and an intercalator, said intercalator being selected from the group consisting of ethidium bromide, actinomycin D, proflavin and acridine orange, and determining the fluorescent polarization of said mixture;
(b) adding the mixture of the compound and the intercalator to a nucleic acid under conditions suitable for the binding of both the compound and the intercalator to the nucleic acid, and determining the fluorescent polarization of the mixture containing the compound, the intercalator and the nucleic acid; and
(c) determining the extent of binding of the compound to the nucleic acid from the difference in the fluorescent polarization of the mixture containing the compound and the intercalator and the mixture containing compound, the intercalator and the nucleic acid.
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Abstract
A novel method for the detection, screening, the quantitation of compounds that interact with nucleic acids (DNA or RNA) is provided. The basis for this method is the inhibition of acridine orange binding to the nucleic acid. Measurement of binding inhibition utilizes the rapid, repeatable technique of fluorescence polarization.
78 Citations
4 Claims
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1. A process for the quantitative determination of the extent of binding of a compound to a nucleic acid, which comprises the steps:
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(a) preparing a mixture containing said compound and an intercalator, said intercalator being selected from the group consisting of ethidium bromide, actinomycin D, proflavin and acridine orange, and determining the fluorescent polarization of said mixture; (b) adding the mixture of the compound and the intercalator to a nucleic acid under conditions suitable for the binding of both the compound and the intercalator to the nucleic acid, and determining the fluorescent polarization of the mixture containing the compound, the intercalator and the nucleic acid; and (c) determining the extent of binding of the compound to the nucleic acid from the difference in the fluorescent polarization of the mixture containing the compound and the intercalator and the mixture containing compound, the intercalator and the nucleic acid. - View Dependent Claims (2, 3, 4)
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Current AssigneeRhone-Poulenc Rorer Pharmaceuticals Incorporated (Sanofi-Aventis U.S., LLC)
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Original AssigneeMeloy Laboratories Incorporated
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InventorsSchulman, Gail E., Richardson, Carol L.
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Primary Examiner(s)Wiseman, Thomas G.
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Application NumberUS06/057,739Time in Patent Office617 DaysField of Search435/6, 435/7, 435/172, 23/230 B, 424/2, 424/7US Class Current436/508CPC Class CodesC12Q 1/68 involving nucleic acidsY10S 436/80 Fluorescent dyes, e.g. rhod...
