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DNA decatenation is an essential step of DNA replication in cells. In bacteria, topoisomerase IV (the parC-parE complex) carries out this reaction, converting concatenated DNA into decatenated DNA.
Kit Components
| Component | GDDA100KE |
|---|---|
| 10 x Buffer | 300 µl |
| Concatenated DNA (kDNA, 60 µg/ml) | 205 µl |
| 10 mM ATP | 205 µl |
| 5 x Gel loading buffer | 550 µl |
| E. coli topoisomerase IV, parC-parE (10 µM) | 22 µl |
Final reaction conditions: 20 mM Tris-HCl pH 8, 35 mM NH4OAc, 4.6% glycerol, 1 mM DTT, 0.005% Brij35, 8 mM MgCl2, 6 µg/ml concatenated DNA, 1 mM ATP and 10 nM E. coli topoisomerase IV. A reaction without enzyme or ATP can serve as negative control.
Assay Principle
Decatenated and concatenated DNA (kDNA) are separated by agarose gel electrophoresis, so E. coli topoisomerase IV activity is seen as the release of decatenated DNA from the concatenated network after staining.
Specifications
| Assay Target | E. coli Topo IV DNA decatenation |
|---|---|
| Target Type | DNA Topoisomerase IV |
| Organism | E. coli |
| Assay Type | Enzyme Activity Assay |
| Detection Method | Gel Electrophoresis |
| Instrument Required | Gel electrophoresis + UV imaging |
| Gel Conditions | 1% agarose in 1 x TAE, 100 V for 90 min to 2 hours |
| Size | 100 assays |
| Storage | Please contact us to confirm storage conditions. |
| Shipping | Shipped frozen. |
Protocol
The manufacturer's protocol describes the procedure and reaction volumes.
- Gel-Based E. coli Topoisomerase IV DNA Decatenation Assay (PDF, see Documents): Covers catalog # GDDA100KE.
Safety & Handling
Can’t find the exact format you need? We can help you source or customize this E. coli Topo IV DNA decatenation assay — including larger pack sizes, bulk reagents, bulk enzyme, or a protocol adapted to your samples. Click Talk to a Scientist, email support@biohippo.com, or explore our Research Services. Our team will be in touch shortly.
