GeneArt Gibson Assembly HiFi Master Mix enables DNA assembly and cloning via a technique that allows multiple overlapping DNA fragments to be seamlessly linked in a one-step, 15–60 minute isothermal reaction. DNA fragments of different lengths are uniformly assembled using complementary overlaps between fragments. The inherent flexibility of this approach is suitable for small and large DNA constructs and includes both single and multiple inserts. The resulting products can be used for a variety of downstream applications including transformation, PCR and rolling circle amplification (RCA). The GeneArt Gibson Assembly HiFi Master Mix kit includes master mix, positive control, and water and accommodates the use of your own competent cells.

Features of the GeneArt Gibson Assembly HiFi Master Mix include:
• Simple—seamlessly assemble and clone up to six DNA fragments in a single reaction
• Efficient—high fidelity provides more correct clones than other methods
• Flexible—design guidelines allow assembly into any vector of your choice
• Convenient—available as master mix kits and cloning kits complete with chemical or electrocompetent cells
• Trusted—over 4000 citations in the scientific literature highlighting great success

Seamless assembly of multiple fragments
GeneArt Gibson Assembly HiFi cloning is a simple, one-step process whereby up to six fragments are combined in a proprietary enzymatic mix in order to assemble DNA fragments with shared terminal end homology without leaving any extra sequences or scars behind (seamless). After a 15–60 minute incubation, a portion of the assembly reaction is then transformed into chemically competent or electrocompetent cells, yielding clones that are ready for analysis the next day. The required 20- to 40-base pair end homology is designed into the de novo fragment for synthesis or can be easily engineered by PCR amplification with custom DNA oligos. This special enzyme mix creates a seamless and covalently bound DNA construct providing high efficiency.

Robust method for maximum efficiency
Due to the covalently bound final product, the GeneArt Gibson Assembly HiFi method allows the utilization of chemically competent cells or electrocompetent cells for the highest transformation efficiency, improving the success of finding the right clone, particularly for more challenging constructs. There is no need for restriction enzymes, ligation, or recombination sites, and the method provides a perfectly seamless construct without unwanted extra bases.

Provides great versatility
The GeneArt Gibson Assembly HiFi method provides versatility, can streamline many techniques through the rapid combination, addition, deletion, or exchange of DNA segments, and eliminates the need to subclone, saving time and effort in the cloning workflow.

Convenient formats
GeneArt Gibson Assembly HiFi kits are available in multiple formats, including:
• GeneArt Gibson Assembly HiFi Cloning Kit, chemically competent cells (with One Shot TOP10 chemically competent cells)
• GeneArt Gibson Assembly HiFi Cloning Kit, electrocompetent cells (with ElectroMAX DH10B electrocompetent cells)
• GeneArt Gibson Assembly HiFi Master Mix kits (cloning kit without the cells) (this page)