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| Product Name | Fast Restriction Enzyme BsaI (Recognition Site: GGTCTC) |
| Catalog No. | IVDRM-0017 |
| Description | This fast restriction enzyme is a recombinant Type IIS endonuclease engineered for rapid, precise DNA cleavage, completing digestion of plasmid DNA, PCR products, and genomic DNA within 5-15 minutes. All enzymes in this fast digestion series share a single universal reaction buffer, which greatly simplifies reaction setup when multiple enzymes are used together. With generous activity overcapacity, the enzyme readily handles excess substrate and difficult templates. Its cleavage pattern makes it well suited for Golden Gate assembly, construct cloning, and routine digestion of plasmids and PCR amplicons. |
| Intended Use | Rapid restriction digestion of plasmid DNA, PCR products, and genomic DNA for molecular cloning workflows, including Golden Gate assembly and construct verification. |
| Principle / Technology | Recombinant Type IIS restriction endonuclease produced by genetic engineering; recognizes an asymmetric sequence and cleaves DNA at a fixed distance outside the recognition site, enabling seamless cloning strategies such as Golden Gate assembly. |
| Sample Type | Plasmid DNA, PCR products, genomic DNA |
| Performance Range / Specifications | Complete digestion achieved in 5-15 minutes at 37°C; enzyme activity of 20 U/μL with high activity redundancy, tolerating substrate excess and difficult templates. |
| Specificity | Specifically recognizes 5'-GGTCTC(N)1-3' and cleaves downstream of the recognition site. Cleavage is blocked when CpG methylation fully overlaps the recognition sequence; Dam, Dcm, EcoKI, and EcoBI methylation have no effect on activity. |
| Reaction Conditions / Protocol | Assemble the reaction in 1× universal digestion buffer and incubate at 37°C for 5-15 min. Heat inactivate at 80°C for 20 min when enzyme removal is required. |
| Components / Formulation | Fast Restriction Enzyme BsaI (20 U/μL) Universal digestion buffer (shared across the fast enzyme series) |
| Storage Conditions | Store at -25°C to -15°C |
| Shelf Life | 2 years |
| Package Specifications | 50 reactions (50 T) |
| Concentration | 20 U/μL |
| Source / Origin | Recombinant enzyme produced by genetic engineering (heterologous expression) |
| Expiration Date / Stability | Stable for 2 years when stored at -25°C to -15°C. |
| Compatibility | Isoschizomers: PalAI, SgsI. Compatible with Golden Gate assembly, cloning construction, and digestion of plasmid and PCR-derived DNA. |
| Recommended Buffer System | 1× universal digestion buffer shared by all enzymes in the fast restriction enzyme series, simplifying multi-enzyme reaction setups. |
| Application Notes / Precautions | Optimal incubation temperature is 37°C; digestion is typically complete within 5-15 min. A single universal buffer is used for all enzymes in this series, streamlining double or multiple digests. High activity overcapacity allows efficient digestion of excess substrate or difficult templates. If downstream applications are sensitive to residual enzyme, heat inactivate at 80°C for 20 min. |
| Recognition Site | 5'-GGTCTC(N)1↓-3' 3'-CCAGAG(N)5↑-5' |
| Heat Inactivation | 80°C for 20 min |
| Methylation Sensitivity | Dam, Dcm, EcoKI, EcoBI: no effect; CpG: cleavage blocked when the methylation site fully overlaps the recognition sequence. |
For research use only, not for clinical use.
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