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At the DNA replication fork, DNA helicase (DnaB) opens double-stranded DNA and bacterial DNA primase (DnaG) builds the RNA primers. Helicases greatly boost primase activity in gram-negative bacteria such as E. coli and H. influenzae, but not in gram-positive S. aureus and S. pneumoniae.
Kit vs Kit Plus
The only difference is the enzyme: PGA100K supplies the assay reagents without enzyme, while PGA100KE adds 45 µl of 100 x S. pneumoniae primase. Both cover 100 assays in a 96-well plate or 200 assays in a 384-well plate.
| Kit (enzyme not included), 100 assays (PGA100K) | 100 assays (96-well) or 200 assays (384-well) |
|---|---|
| Kit Plus (enzyme included), 100 assays (PGA100KE) | 100 assays (96-well) or 200 assays (384-well) |
Kit Components
| Component | Kit (enzyme not included), 100 assays (PGA100K) | Kit Plus (enzyme included), 100 assays (PGA100KE) |
|---|---|---|
| 10 x Assay buffer | 600 µl | 600 µl |
| 100 x DNA template | 45 µl | 45 µl |
| 100 x NTP mix | 45 µl | 45 µl |
| 10 x Fluorescence dye | 850 µl | 850 µl |
| 100 x S. pneumoniae primase | Not included | 45 µl |
Assay solution: 10 mM MES pH 6.5, 5 mM magnesium acetate, 12.5 mM ammonium chloride, 0.5 mM DTT, 5 mM calcium chloride, 0.003% Brij-35, 100 nM DNA, 0.5 mM NTPs, 100 nM enzyme.
Assay Principle
The assay measures the RNA primers made by DNA primase from a DNA template and NTPs. After the reaction a fluorescence dye is added and the signal is read at 535 nm (excitation 485 nm).
Specifications
| Assay Target | S. pneumoniae DNA primase activity |
|---|---|
| Target Type | DNA Primase |
| Organism | S. pneumoniae |
| Assay Type | Enzyme Activity Assay; Inhibitor Screening Assay |
| Detection Method | Fluorescence |
| Detection Wavelength | Ex 485 nm / Em 535 nm |
| Plate Format | 96-well or 384-well plate |
| Instrument Required | Fluorescence microplate reader |
| Assay Time | About 65 min (60 min reaction + 5 min detection) |
| Reference | Lacriola CJ et al., The Journal of Antibiotics 70, 918-920 (2017) |
| Size | 100 assays (96-well) or 200 assays (384-well) |
| 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 Primase Assay (PDF, see Documents): Covers catalog # PGA100K, PGA100KE.
Safety & Handling
Can’t find the exact format you need? We can help you source or customize this S. pneumoniae DNA primase 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.
