Generate seamless plasmid DNA or BAC clones without restriction digestion

The Gibson Assembly process begins by designing dsDNA fragments with 20 – 40bp overlapping ends. For the Gibson Assembly HiFi 1-Step reaction, these DNA fragments are combined with a 2X Gibson Assembly® HiFi 1-Step Master Mix and incubated. This during this time, complementary ends are generated, annealed, and sealed. This this one-step, isothermal process can be performed in less than 60 minutes and results in a transfection or transformation ready, double stranded, fully ligated DNA construct.

The GA 1-Step Master Mix (2X) contains a proprietary mixture of enzymes and reagents optimized to facilitate one-step assembly of double stranded DNA fragments. This mastermix includes a proof-reading polymerase that mediates junction repair resulting assembled constructs with low rates of junction errors and high sequence fidelity. This product does not include the GA Positive Control found in the Gibson Assembly® HiFi 1-Step Kit. If you are new to Gibson Assembly or require a validated positive control for you experiments, you may want to consider catalog numbers GA1100-S, GA1100-10, or GA1100-50.

The Gibson Assembly method allows the insertion of one or more linear double stranded DNA fragments into a virtually any vector without the need to rely on compatible restriction sites. Gibson Assembly is significantly faster than traditional restriction enzyme digest-based cloning and proven for the cloning of both small and large double stranded DNA fragments. Using the Gibson Assembly® HiFi 1-Step Master Mix, the simultaneous cloning of one to five inserts with sizes ranging from 500 bp to 32kb can be accomplished in a single tube, in a single step, using a robust isothermal reaction that yields cloning efficiencies of greater than 90%.