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DNA topoisomerase II – DNA supercoiling assay
DNA topoisomerases such as bacterial topoisomerase II (gyrase) convert relaxed circular DNA into supercoiled DNA. The DNA Topoisomerase II (Gyrase) Assay Kit is based on the principle that the supercoiled DNA and relaxed DNA yield different fluorescent intensity when interact with fluorescence dye H19. The relaxed DNA suppresses the fluorescent intensity much more than the supercoiled DNA. Therefore, when the relaxed DNA is converted into its supercoiled form, the fluorescent signal increases. The change of fluorescence intensity is used to measure the supercoiling reaction.
Each kit (Catalog number DSA020K) includes the reaction buffer (T2 Buffer), relaxed plasmid DNA and fluorescence dye H19 for 20 assays of DNA supercoiling reactions a 96-well plate or 40 assays in a 384-well assay plate. The reaction buffer is optimized for bacterial topoisomerase II (gyrase). Bacterial gyrases are provided separately.
Each kit (Catalog number DSA020KE) includes the reaction buffer (T2 Buffer), relaxed plasmid DNA, E .coli gyrase and fluorescence dye H19 for 20 assays of DNA supercoiling reactions a 96-well plate or 40 assays in a 384-well assay plate. The reaction buffer is optimized for E. coli topoisomerase II (gyrase).
The Reagent Set (Catalog No. DSA1000) is designed for drug discovery HTS uses. Each reagent set includes the reaction buffer and fluorescence dye H19 for 1000 assays of DNA supercoiling reactions a 96-well plate assay format or 2000 assays in a 384-well assay format. Relaxed DNA and gyrase are not included in the set. Gyrase is provided separately. Relaxed plasmid pBR322 from any commercial source can be used.
Wild-type E. coli topoisomerase II (gyrase) for 100 assays.
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