![]() ![]() There is no need for restriction enzymes, ligation, or recombination sites, and the method provides a perfectly seamless construct without unwanted extra bases. This special enzyme mix creates a seamless and covalently bound DNA construct providing high efficiency.ĭue 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. 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. 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. 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). ![]() Trusted-over 4000 citations in the scientific literature highlighting great success.Convenient-available as master mix kits and cloning kits complete with chemical or electrocompetent cells.Flexible-design guidelines allow assembly into any vector of your choice.Efficient-high fidelity provides more correct clones than other methods.Simple-seamlessly assemble and clone up to six DNA fragments in a single reaction.coli.įeatures of the GeneArt Gibson Assembly HiFi Cloning Kit include: The GeneArt Gibson Assembly HiFi Cloning Kit, electrocompetent cells, is a complete kit that includes master mix, positive control, water, and One Shot TOP10 chemically competent E. The resulting products can be used for a variety of downstream applications including transformation, PCR and rolling circle amplification (RCA). The inherent flexibility of this approach is suitable for small and large DNA constructs and includes both single and multiple inserts. DNA fragments of different lengths are uniformly assembled using complementary overlaps between fragments. The GeneArt Gibson Assembly HiFi Cloning Kit 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.
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