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| Main SKU |
DNA polymerase III synthesizes DNA at the bacterial replication fork, extending the RNA primer made by DNA primase. The alpha subunit is the catalytic subunit of the polymerase, and the kit can be used both to detect bacterial DNA polymerase III activity and for high-throughput screening of bacterial DNA polymerase inhibitors.
Kit Components
| Component | DPA100KN |
|---|---|
| 10 x Buffer DP | 500 µl |
| 100 x DNA template | 35 µl |
| 100 x dNTP mix | 35 µl |
| 10 x Fluorescence dye | 320 µl |
| 100 x S. pneumoniae DNA polymerase III alpha subunit | 35 µl |
Final reaction conditions: 20 mM HEPES pH 7.5, 10 mM Mg(OAc)2, 1 mM DTT, 30 nM DNA, 20 nM DNA polymerase III alpha subunit, 0.1 mM dATP and 0.1 mM dGTP.
Assay Principle
The kit measures the DNA synthesized by the polymerase from a DNA template and dNTPs. After the reaction a fluorescence dye is added and the signal is read at 535 nm (excitation 485 nm); higher polymerase activity gives more signal.
Specifications
| Assay Target | S. pneumoniae Pol III alpha activity |
|---|---|
| Target Type | DNA Polymerase |
| Organism | S. pneumoniae |
| Assay Type | Enzyme Activity Assay; Inhibitor Screening Assay |
| Detection Method | Fluorescence |
| Detection Wavelength | Ex 485 nm / Em 535 nm |
| Plate Format | 384-well or 96-well plate |
| Instrument Required | Fluorescence microplate reader |
| Assay Time | About 65 min (60 min reaction + 5 min detection) |
| 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.
- S. pneumoniae DNA Polymerase Assay (PDF, see Documents): Covers catalog # DPA100KN.
Safety & Handling
Can’t find the exact format you need? We can help you source or customize this S. pneumoniae Pol III alpha activity 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.