SGI-DNA’s Gibson Assembly® HiFi master mix is the original Gibson Assembly® cloning method, which has been optimized for the scarless joining of up to five inserts into a plasmid vector, resulting in DNA constructs with reliably low error rates. This single-tube, isothermal reaction is compatible with fragments ranging from 500 bp to 32 kb and can be completed in 60 minutes or less. Unlike other approaches, SGI-DNA’s Gibson Assembly® process utilizes Phusion® high-fidelity DNA polymerase for 3’ end extension. This proof-reading process results in higher numbers of error-free clones ready for multiple downstream applications.
For less concentrated DNA samples, the 4X Gibson Assembly® HiFi HC 1-step kit is ideal, requiring only 2.5 µL of mix per 10 µL reaction. Both the original 2X master mix and 4X high concentration (HC) master mix can assemble multiple DNA fragments with greater than 90% cloning efficiency.
There are over 4,000 citations in the scientific literature on the Gibson Assembly® cloning method: Gibson et al. 2009. Nature Methods.