FAQ: Which restriction enzymes are used in GoldenBraid Assembly?

BsaI is one Type IIS enzyme used in this method. NEB also offers an engineered high-fidelity version of this enzyme, BsaI-HF, which has the added benefit of being Time-Saver™ qualified (can digest DNA in 5-15 mins and can be digested safely overnight) and exhibits reduced star activity. However methylation sensitivities differ between the two enzymes. BsaI-HF works in CutSmart® Buffer, as does T4 DNA Ligase (also part of the GoldenBraid workflow) which is 100% functionally active in this buffer, when the reaction is supplemented with 1 mM ATP. Other Type IIS enzymes used in GoldenBraid include BsmBI, BtgZI and BbsI/BbsI-HF.


GoldenBraid 2.0: A Comprehensive DNA Assembly Framework for Plant Synthetic Biology

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