Invitrogen™

GeneArt™ Gibson Assembly EX Cloning Kit, chemically competent cells

Catalog number:  A46633

Related applications:

Seamless Cloning & Genetic Assembly

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Description

The GeneArt Gibson Assembly EX Cloning Kit enables DNA assembly via a technique that allows up to 15 overlapping DNA fragments to be seamlessly linked in a single-tube, two-step optimized reaction. DNA fragments of different lengths are uniformly assembled using complementary overlaps between fragments. The inherent flexibility and success of this approach is suitable for small and large DNA constructs, includes both single and multiple inserts, and has been used to build entire genomes. The resulting products can be used for a variety of downstream applications, including transformation, PCR, and rolling circle amplification (RCA). The GeneArt Gibson Assembly EX Cloning Kit, chemically competent cells, is a complete kit that includes master mix, positive control, water, and One Shot TOP10 chemically competent E. coli.

Features of the GeneArt Gibson Assembly EX Cloning Kit include:
Simple—seamlessly assemble and clone up to 15 DNA fragments in a single reaction
Efficient—robust efficiency provides successful clones for both simple and challenging constructs
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 up to 15 fragments
GeneArt Gibson Assembly EX cloning is a robust, single-tube, two-step process whereby up to 15 inserts and vector 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 this dually optimized reaction is complete, a portion of the assembly reaction is then transformed into chemically competent or electrocompetent cells, yielding clones 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 EX method allows the utilization of chemically competent cells or electrocompetent cells for the highest transformation efficiency, especially important for 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 EX 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 EX kits are available in multiple formats, including:
• GeneArt Gibson Assembly EX Cloning Kit, chemically competent cells (with One Shot TOP10 chemically competent cells) (this page)
GeneArt Gibson Assembly EX Cloning Kit, electrocompetent cells (with ElectroMAX DH10B electrocompetent cells)
GeneArt Gibson Assembly EX Master Mix kits (cloning kit without the cells)

For Research Use Only. Not for use in diagnostic procedures.

Specifications

Number of Fragments: Up to 15 fragments
Shipping Condition: Dry Ice
Workflow Step: DNA Assembly
Product Line: GENEART™

Contents & storage

Includes master mix, positive control, water, and One Shot TOP10 chemically competent E. coli.