Attention Golden Gate Assembly users: BsaI-HFv2 (NEB #R3733) has been optimized for Golden Gate Assembly*. BsaI-HFv2 also works well for any protocol requiring DNA cutting by BsaI. This is the recommended enzyme for any purpose requiring digestion at the recognition sequence ...5′-GGTCTC(N1)/(N5)-3′...
*The requirements for digestion of DNA during Golden Gate Assembly are more demanding than what is encountered in traditional DNA cloning. For Golden Gate, the enzyme must function well in a buffer that might not be the same buffer as that normally provided for the restriction enzyme, and maintain activity for an extended time at elevated temperatures in a dynamic cutting/re-ligating reaction with competition for substrate binding between the endonuclease and ligase. Extensive testing has demonstrated superior performance of BsaI-HFv2 compared to both BsaI (NEB #R0535) and BsaI-HF (NEB #R3535) in this challenging Golden Gate assembly context where restriction enzyme efficiency and fidelity are critically important.
- Engineered for improved performance
- Used in Golden Gate Assembly; maintains high activity levels in T4 DNA Ligase Buffer often used in this application
- Type IIS restriction enzymes recognize asymmetric DNA sequences and cleave outside of their recognition sequence
- 100% activity in CutSmart Buffer
- Time-Saver™ qualified for digestion in 5-15 minutes
- Reduced star activity
"I really, really love BsaI-HFv2. We are back to getting phenomenal GGA results again with very little background."
-M.C, Principal Investigator, Davidson College, NC
Restriction Enzymes in Golden Gate Assembly
Golden Gate Assembly Workflow
Product SourceAn E. coli strain that carries the cloned and modified BsaI gene from Bacillus stearothermophilus 6-55 (Z. Chen)
The following reagents are supplied with this product:
Store at (°C) Concentration Gel Loading Dye, Purple (6X) 25 6 X CutSmart® Buffer -20 10 X
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