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| Product Name | DNA Rapid Isothermal Amplification Kit (Basic) |
| Catalog No. | RESK-0118 |
| Description | A raw-material reagent kit for recombinase polymerase amplification (RPA), developed with recombinase, single-stranded DNA binding protein, and strand-displacing DNA polymerase for isothermal amplification of DNA nucleic acid targets. Only a pair of specific primers and template need to be added to the reaction system for isothermal amplification; the purified amplification product is then analyzed by agarose gel electrophoresis for result interpretation. In RPA, recombinase proteins form complexes with primers that search for homologous sequences on double-stranded DNA. The recombinase strand-exchange activity inserts the primer at the homologous site, and single-stranded binding proteins stabilize the displaced DNA strand. The recombinase is then disassembled so that the primer 3' end is strand-displaced and bound by DNA polymerase, which extends the primer; repeating this cycle exponentially amplifies the target region of the template. |
| Intended Use | For isothermal amplification and rapid detection of DNA targets in research and assay development; widely applicable to in vitro diagnostics, veterinary testing, food safety, biosafety, and agriculture. |
| Principle / Technology | Recombinase polymerase amplification (RPA): recombinase proteins form complexes with primers that search for homologous sequences on double-stranded DNA; the recombinase strand-exchange activity inserts the primer at the homologous site while single-stranded binding proteins stabilize the displaced DNA strand. The recombinase is then disassembled, leaving the primer 3' end accessible to a strand-displacing DNA polymerase that extends the primer; repeating this cycle exponentially amplifies the target region. Multiple enzymes participate, including T4 UvsX, T4 UvsY, GP32, Bsu DNA polymerase, and creatine kinase (CK). |
| Detection Method | Agarose gel electrophoresis of the purified amplification product; positive samples show a clear, single band, while negative samples show no obvious band. |
| Sample Type | DNA nucleic acid templates. |
| Performance Range / Specifications | Multi-enzyme system involving T4 UvsX, T4 UvsY, GP32, Bsu DNA polymerase, and CK. Isothermal reaction at 37-42 °C for 10-30 min amplifies trace nucleic acid to a detectable level. Detection sensitivity down to a single copy or tens of copies (or fewer) of DNA. Low instrument dependence; simple operation and straightforward result interpretation. Widely used in in vitro diagnostics, veterinary, food safety, biosafety, and agriculture. Successfully amplifies targets with GC contents of 40%-70% and lengths of 100-500 bp, yielding a clear, unique band for positives and no obvious band for negatives. |
| Sensitivity / LOD | Down to a single copy or tens of copies (or fewer) of DNA; demonstrated superior detection sensitivity in gradient-dilution testing of positive control templates. |
| Specificity | Sequence-specific amplification driven by a pair of specific primers; positive targets of varying GC content (40%-70%) and length (100-500 bp) produce a clear, single electrophoresis band, while negatives show no obvious band. |
| Reaction Conditions / Protocol | Isothermal incubation at 37-42 °C for 10-30 min. Add a pair of specific primers and template to the reaction system; after amplification, purify the product and analyze by agarose gel electrophoresis. |
| Components / Formulation | RPA enzyme system including T4 UvsX recombinase, T4 UvsY, GP32 single-stranded DNA binding protein, Bsu DNA polymerase, and creatine kinase (CK). Primers and template are supplied by the user. |
| Storage Conditions | Store at -25 to -15 °C. |
| Shelf Life | 1 year. |
| Package Specifications | 16 T; 48 T; 100 T |
| Product Form | Liquid reagent kit (isothermal amplification raw-material premix). |
| Expiration Date / Stability | Valid for 1 year when stored at -25 to -15 °C. |
| Compatibility | Suitable for DNA targets with GC contents of 40%-70% and amplicon lengths of 100-500 bp. |
| Application Notes / Precautions | Purify the amplification product before agarose gel electrophoresis for result interpretation. Use a pair of specific primers designed for the target of interest. |
For research use only, not for clinical use.
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