METHOD AND KIT FOR IDENTIFICATION OF NUCLEIC ACID MODIFICATION ENZYMES AND INHIBITORS THEREOF

摘要

A method and kit for detecting nucleic acid modification enzyme activity by monitoring the change in fluorescence polarization upon the change in mass of a fluorescently labeled oligo. A nucleic acid substrate comprises a template having a nucleic acid sequence annealed to a labeled oligo, wherein there is a nick or break located between the adjacent ends of the template and the labeled oligo. A nucleic acid modification enzyme is added to the substrate to form a reaction mixture. A stop reagent may or may not be used to further process the reaction mixture. A test compound is added to the reaction mixture, preferably prior to addition of the nucleic acid modification enzyme, whereby the resulting fluorescence polarization signal is compared to a standard signal to determine if the test compound inhibits nucleic acid modification enzyme activity.