Description: DNA topoisomerase fluorescence assay kit is designed for assaying DNA activities of a DNA topoisomerase that relaxes supercoiled plasmid pAB1 and also for drug discovery research targeting DNA topoisomerases. This kit includes enough enzymes and other reagents for 100 assays: E. coli DNA topoisomerase I (200 units, Cat. No. TB-E-003) and 10xDNA topoisomerase I assay buffer (Cat. No. TB-B-005), DNA topoisomerase I dilution buffer (Cat. No. TB-B-006), T5 exonuclease (Cat. No. TB-E-002, 200 uL of 5 uM) and 10xT5 exonuclease assay buffer (Cat. No. TB-B-003), supercoiled pAB1 (100 uL of 0.2 mg/mL supercoiled pAB1, Cat. No. TB-D-002), and 40xSYBR Green. Figure 1 is the strategy of this DNA topoisomerase relaxation assay. Please see reference 2 for details. For other DNA topoisomerases except DNA gyrase, please use a buffer for the specific topoisomerase. E. coli DNA topoisomerase I can be used as a control.
This kit works for all DNA topoisomerases except bacterial DNA gyrase.
Shipping and storage: DNA topoisomerase fluorescence assay kit is shipped and should be stored in a -80°C freezer (-70-80°C) and is stable for at least 12 months in this condition. E. coli DNA topoisomerase I and T5 exonuclease can be aliquoted on first thawing to prevent loss of activities from freeze-thaw cycles.
This kit works for all DNA topoisomerases except bacterial DNA gyrase.
Shipping and storage: DNA topoisomerase fluorescence assay kit is shipped and should be stored in a -80°C freezer (-70-80°C) and is stable for at least 12 months in this condition. E. coli DNA topoisomerase I and T5 exonuclease can be aliquoted on first thawing to prevent loss of activities from freeze-thaw cycles.
This product is used for research only.
Topoisomerase Fluorescence DNA Relaxation Assay Instruction: