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| Product Name | Restriction Enzyme BbvCI (Recognition Site: 5'-CC↓TCAGC-3') |
| Catalog No. | IVDRM-0150 |
| Description | BbvCI is a Type IIA restriction endonuclease produced by recombinant expression of the BbvCI gene from Bacillus brevis (L. Ge) in Escherichia coli. The enzyme exists as a heterodimer composed of two distinct catalytic subunits, each with its own independent cleavage site. BbvCI recognizes and cuts the non-palindromic sequence CCTCAGC(-5/-2), generating sticky ends with a 3-nucleotide 5' overhang. The enzyme is supplied with a universal reaction buffer that supports double digests with other enzymes in the same buffer system. Prolonged digestion (over 3 hours) is not recommended, to avoid star activity. |
| Intended Use | Molecular cloning, restriction digestion, and double-digest workflows with compatible enzymes in the universal buffer system. |
| Principle / Technology | BbvCI is a Type IIA restriction endonuclease that functions as a heterodimer of two distinct catalytic subunits, each responsible for cleaving one DNA strand. It recognizes the non-palindromic sequence 5'-CCTCAGC-3' and cleaves at positions -5/-2 relative to the site, producing a 3-nucleotide 5' overhang. |
| Sample Type | DNA substrates such as lambda DNA (λDNA) bearing BbvCI recognition sites. |
| Performance Range / Specifications | Concentration: 2 U/μL. Unit definition: One unit (U) is the amount of enzyme required to completely digest 1 µg of λDNA in 1 hour at 37°C in a 50 µL reaction. Cleavage generates sticky ends with a 3-nucleotide 5' overhang. Prolonged digestion (over 3 hours) may cause star activity and is not recommended. CpG methylation of the recognition site affects cleavage. |
| Specificity | Specifically recognizes the non-palindromic sequence 5'-CC↓TCAGC-3' (complementary strand 3'-GGAGT↑CG-5'), cutting at CCTCAGC(-5/-2). |
| Reaction Conditions / Protocol | Optimal reaction: incubate at 37°C in 1× universal reaction buffer. Avoid digestion times longer than 3 hours. Heat inactivation: 80°C for 20 min. |
| Components / Formulation | BbvCI restriction enzyme (2 U/μL) and 1× universal reaction buffer. |
| Storage Conditions | Store at -25°C to -15°C. |
| Shelf Life | Valid for 2 years when stored at -25°C to -15°C. |
| Package Specifications | 50 U |
| Concentration | 2 U/μL |
| Activity / Unit Definition | One unit (U) is defined as the amount of enzyme required to completely digest 1 µg of λDNA in 1 hour at 37°C in a 50 µL reaction system. |
| Source / Origin | BbvCI gene from Bacillus brevis (L. Ge), recombinantly expressed in Escherichia coli. |
| Expiration Date / Stability | Stable for 2 years from the date of manufacture when stored at -25°C to -15°C. |
| Compatibility | Supplied with a universal reaction buffer that allows double digestion with other restriction enzymes in the same buffer system. |
| Recommended Buffer System | 1× universal reaction buffer (supplied). |
| Application Notes / Precautions | Do not extend digestion beyond 3 hours to avoid star activity. The universal buffer enables convenient double digests with other enzymes in the same buffer system. Heat-inactivate at 80°C for 20 min before downstream applications. |
| Recognition Site | 5'-CC↓TCAGC-3' 3'-GGAGT↑CG-5' |
| Heat Inactivation | 80°C for 20 min. |
| Methylation Sensitivity | CpG methylation: cleavage is affected. |
For research use only, not for clinical use.
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