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| Product Name | Restriction Enzyme BpiI (Recognition Site: 5'-GAAGAC(N)2↓-3') |
| Catalog No. | IVDRM-0151 |
| Description | BpiI is a Type IIS restriction endonuclease that specifically recognizes the sequence GAAGAC and cuts downstream of the site, generating 4-nucleotide 5' overhangs. It is one of the most commonly used enzymes for Golden Gate assembly. Supplied with a universal reaction buffer, BpiI can be combined with other restriction enzymes of the same buffer system for double digests, making it equally suitable for routine cloning and restriction analysis beyond Golden Gate applications. |
| Intended Use | Golden Gate assembly, as well as routine molecular cloning and restriction analysis (digestion-based verification). |
| Principle / Technology | BpiI is a Type IIS restriction endonuclease that recognizes the asymmetric sequence 5'-GAAGAC-3' and cleaves the DNA at fixed positions downstream of the recognition site, generating 4-nucleotide 5' overhangs. Because cleavage occurs outside the recognition sequence, the recognition site is removed from the final product, enabling seamless Golden Gate assembly. |
| Sample Type | DNA substrates such as lambda DNA (λDNA) bearing BpiI recognition sites. |
| Performance Range / Specifications | Concentration: 10 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 produces 4-nucleotide 5' overhangs. |
| Specificity | Specifically recognizes 5'-GAAGAC(N)2↓-3' (complementary strand 3'-CTTCTG(N)6↑-5') and cleaves downstream of the recognition sequence. |
| Reaction Conditions / Protocol | Optimal reaction: incubate at 37°C in 1× universal reaction buffer. Heat inactivation: 80°C for 20 min. |
| Components / Formulation | BpiI restriction enzyme (10 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 | 250 U |
| Concentration | 10 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. |
| Expiration Date / Stability | Stable for 2 years from the date of manufacture when stored at -25°C to -15°C. |
| Compatibility | Isoschizomers: BstV2I, BbsI. Compatible with other restriction enzymes in the universal buffer system for double digestion. |
| Recommended Buffer System | 1× universal reaction buffer (supplied). |
| Application Notes / Precautions | BpiI cuts downstream of its recognition site, so the recognition sequence is absent from Golden Gate assembly products, enabling seamless cloning. The universal buffer supports double digests with other enzymes in the same buffer system. Heat-inactivate at 80°C for 20 min when residual activity must be removed. |
| Recognition Site | 5'-GAAGAC(N)2↓-3' 3'-CTTCTG(N)6↑-5' |
| Heat Inactivation | 80°C for 20 min. |
For research use only, not for clinical use.
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