GeneArt™ Gibson Assembly HiFi Master Mix

GeneArt Gibson Assembly HiFi Master Mix enables seamless, one-step, single-tube assembly of multiple overlapping DNA fragments in an isothermal reaction. This flexible DNA assembly methodology works for both small and large constructs, accommodating single or multiple inserts. PCR amplified fragments can be assembled without purification and transformed following a rapid competent cell transformation protocol. This approach significantly reduces hands-on time without great losses in positive clone yields. The resulting products are suitable for various downstream applications, including competent cell transformation, PCR, rolling circle amplification (RCA), synthetic biology, in vitro cell-free workflows, and mutagenesis.

Features of the GeneArt Gibson Assembly HiFi Master Mix include:

• One-step, single-tube reaction with scarless fragment assembly—simplifies the cloning process by using complementary overlaps to join DNA fragments without the need of restriction enzymes and ligases
• High accuracy and fidelity—precise assembly with low rates of junction errors, providing high sequence fidelity
• Multiple insert assembly for a wide range of fragment sizes—capable of assembling up to five DNA fragments plus a vector in one reaction, suitable for assembling DNA fragments ranging from 0.5 to 32 kb
• Trusted and widely cited—over 4000 citations in the scientific literature, highlighting its great success and reliability

Seamless assembly of multiple fragments

GeneArt Gibson Assembly HiFi Master Mix, with its proprietary enzyme blend, is used to seamlessly assemble DNA fragments with shared terminal end homology without leaving any extra sequences behind. The required 20- to 40-base pair terminal end overlaps are 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. This special enzyme mix creates a seamless and covalently bound DNA construct with high efficiency.

Provides great versatility

GeneArt Gibson Assembly HiFi has emerged as a powerful molecular cloning method that enables researchers to engineer constructs that would be too cumbersome or impossible using traditional techniques. This versatile technique can be used in various genetic engineering, synthetic biology, and genome editing applications. Technical notes are provided for site-directed mutagenesis cell-free protein expression workflow combined with RCA.

Convenient formats

GeneArt Gibson Assembly HiFi kits are available in multiple formats, including:

• GeneArt Gibson Assembly HiFi Cloning Kit, chemically competent cells (Cat. No. A46624 with One Shot TOP10 chemically competent cells)
• GeneArt Gibson Assembly HiFi Cloning Kit, electrocompetent cells (Cat. No. A46626 with ElectroMAX DH10B electrocompetent cells)
• GeneArt Gibson Assembly HiFi Master Mix kits (cloning kit without the cells) (this page)

For more challenging DNA assembly applications such as cloning large or up to 15 fragments simultaneously, GeneArt Gibson Assembly EX should be considered (Cat. No. A46633, A46634, A46635, A46636, A46637).