Gibson Assembly Kits

• Gibson 1-Step Assembly of 1 to 5 Inserts
• Gibson 2-Step Assembly of 6 to 15 Inserts
• Gibson Assembly Site-Directed Mutagenesis

The Gibson Assembly® Method Overview
Gibson Assembly® cloning allows for insertion of DNA fragments into virtually any vector without the need for compatible restriction sites. Join any two fragments to generate seamless constructs in your desired vector.

• Significantly faster than traditional cloning methods
• Avoid time-consuming subcloning
• Insert 1-15 fragments in a single round of cloning in ~1 hour
• Achieve cloning efficiencies of up to 95%
• E. cloni 10G Chemically Competent Cells ideal for Gibson Assembly, see link below

The Gibson Assembly® method can be used to rapidly clone multiple DNA fragments into any vector in one hour or less without the use of restriction enzymes. By designing DNA fragments with homologous overlapping ends, users of the Gibson Assembly® method can create DNA constructs in a single round of cloning.
The method is initiated by combining DNA fragments with the Gibson Assembly® Master Mix. The master mix enzyme cocktail mediates strand chew back, exposing a single strand which allows for annealing of the terminal homologous overlap sequences.
Annealing of the homologous overlap sequences is followed by extension and ligation to yield an assembled product. Seamless assembly can be readily applied to both routine cloning and large and complex cloning projects.

The Gibson Assembly® HiFi 1-Step Kit allows for the assembly of up to 5 different fragments ranging from 500 bp to 32 kb using an isothermal process. As implied by the name, the HiFi 1-Step process is performed in a single step.

The Gibson Assembly® Ultra Kit is optimal for more complex constructs (up to 15 fragments) or for constructs incorporating fragments from 100 bp to 100 kb.

The Gibson Assembly® Site-Directed Mutagenesis (SDM) Kit enables the incorporation of multiple targeted mutations into a construct. Using this single kit solution, it is possible to add any combination of substitution, insertion, or deletion mutations in a single round of mutagenesis and cloning.

References
Gibson DG et al. (2009) Enzymatic assembly of DNA molecules up to several hundred
kilobases. Nature Methods 6(5):343-5.
Gibson DG et al. (2010) Creation of a bacterial cell controlled by a chemically synthesized genome. Science 329(5987):52-6.

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Related Links

Gene Fragment Synthesis Service
Gibson Assembly Cloning Guide
The Gibson Assembly® Primer Design Tool
E. cloni 10G and Gibson Assembly Application Note
E. cloni 10G Competent Cells