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| Product Name | Nicking Endonuclease Nb.BbvCI (Recognition Site: 5'-CCTCAGC-3', Bottom-Strand Nicking) |
| Catalog No. | IVDRM-0162 |
| Description | Nb.BbvCI is a nicking endonuclease that recognizes the sequence 5'-CCTCAGC-3' and cleaves only one strand (the bottom strand) of a double-stranded DNA substrate, introducing a single-stranded nick instead of cutting the duplex through. The enzyme is widely used in isothermal nucleic acid amplification methods such as SDA and RCA: the nick generated by Nb.BbvCI primes polymerase-mediated strand displacement, and repeated cycles of nicking, displacement, and extension drive exponential amplification of the target nucleic acid. |
| Intended Use | For molecular cloning and nucleic acid isothermal amplification applications (e.g., SDA, RCA), where enzyme-generated nicks trigger polymerase strand displacement to achieve exponential amplification. |
| Principle / Technology | Nicking endonuclease: cleaves only one strand of the dsDNA substrate. Nb.BbvCI nicks the bottom strand within its recognition site (3'-GGAGT↑CG-5'), producing a nick rather than a double-strand break. |
| Sample Type | Double-stranded DNA (dsDNA) substrates. |
| Performance Range / Specifications | Concentration: 10 U/µL. One unit (U) is the amount of enzyme required to completely convert 1 µg of supercoiled p615 DNA to the open circular form in 1 hour at 37°C in a 50 µL reaction. |
| Specificity | Recognizes 5'-CCTCAGC-3' and nicks the bottom strand. Methylation effect: EcoBI methylation blocks cleavage. |
| Reaction Conditions / Protocol | 1× reaction buffer; incubate at 37°C. Heat inactivation: 80°C for 20 min. |
| Components / Formulation | Nb.BbvCI nicking endonuclease, 10 U/µL; supplied with the recommended reaction buffer. |
| Storage Conditions | Store at -25°C to -15°C. |
| Shelf Life | 2 years when stored at -25°C to -15°C. |
| Package Specifications | 300 U |
| Concentration | 10 U/µL |
| Activity / Unit Definition | One unit (U) is defined as the amount of enzyme required to completely convert 1 µg of supercoiled p615 DNA into the open circular form within 1 hour at 37°C in a 50 µL reaction. |
| Expiration Date / Stability | Stable for 2 years at -25°C to -15°C. |
| Recommended Buffer System | 1× universal restriction enzyme reaction buffer (supplied). |
| Application Notes / Precautions | Incubate reactions at 37°C for optimal activity. EcoBI methylation of the substrate blocks cleavage; use methylation-free DNA when possible. Inactivate the enzyme at 80°C for 20 min before downstream steps. Avoid repeated freeze-thaw cycles; store at -25°C to -15°C. |
| Recognition Site | 5'-CCTCA GC-3' 3'-GGAGT↑CG-5' (Nb.BbvCI cleaves the bottom strand, as indicated by ↑.) |
| Heat Inactivation | 80°C for 20 min. |
| Methylation Sensitivity | EcoBI: cleavage blocked. |
For research use only, not for clinical use.
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