Thermo Fisher Scientific GeneArt High-Order Genetic Assembly Systems, with yeast growth media

Invitrogen™ GeneArt™ High-Order Genetic Assembly Systems (with yeast growth media)

The GeneArt® High-Order Genetic Assembly System is a highly efficient kit for the simultaneous and seamless assembly of up to 10 DNA fragments, totaling up to 110 Kbp in length, into any vector. It utilizes yeast’s natural recombination ability, minimizing in vitro DNA handling and eliminating enzymatic treatments such as restriction and ligation, while enabling precise DNA sequence fusions. The kit includes materials for yeast transformation and purification, yeast selective media, and competent E. coli for plasmid amplification.

주요 특징

• Easy and Powerful — Clone up to 10 DNA fragments (up to 110 Kbp) in a single vector; no restriction digestion, ligation, or recombination sites required
• Precision and Efficiency — Achieve up to 90% correct clones with no extra sequences left behind
• Flexibility — Use the provided linear vector or your own compatible vector; clone fragments without end-homology
• Free Tools — Web-based design interface for oligo design and project planning
• Diverse Applications — Accelerate synthetic and molecular biology workflows through rapid DNA segment manipulation

For cloning 1–4 smaller fragments via an in vitro method, consider the GeneArt® Seamless Cloning and Assembly Kit (Cat. No. A13288).

Easily Create New Specific Constructs

This system uses transformation-associated recombination (TAR) in Saccharomyces cerevisiae to join pre-existing DNA fragments or synthetic oligonucleotides into a single recombinant molecule. Fragments are joined via shared end-terminal homology or synthetic recombination linkers when homology is absent. The process is seamless and efficient, optimized for up to 110 Kbp and 10 fragments.

Simple Clone Verification

Recombinant yeast clones undergo a simplified 10-minute DNA extraction protocol using included lysis buffer and glass beads. The extracted DNA is suitable for colony PCR and direct E. coli transformation for downstream analysis.

Choosing a Cloning Vector

Requires shuttle vectors compatible with yeast and E. coli (e.g., BAC-YAC). No vector is included, but the GeneArt® pYES1L Vector with Sapphire Technology™ (Cat. No. A13287) is available separately. Compatibility can also be achieved using the GeneArt® High-Order Vector Conversion Cassette (Cat. No. A13291).

Cloning Efficiency

Efficiency depends on fragment size, number, and homology. Terminal end types (A-overhangs or blunt) do not affect efficiency.

Typical efficiencies (using GeneArt® pYES1L Vector):

• 90% for 5 fragments of 10 Kb each

• 90% for 10 fragments of 5 Kb each

• 50% for 10 fragments of 10 Kb each

Efficiencies for fragments without end-homology (using stitching oligos):

• 90% for 1 fragment of 10 Kb

• 75% for 2 fragments of 10 Kb each

Design Your Cloning in silico

Free online design tool helps design fragments and oligos with correct homology and spacing. It verifies compatibility, designs oligos for PCR or stitching, and provides a graphical vector map and downloadable annotated GenBank sequence compatible with Vector NTI® software.

Applications

Ideal for molecular and synthetic biology experiments including:

• Modular vector construction with interchangeable parts
• Fusion protein creation
• DNA element deletion, replacement, or addition
• Cloning large fragments without end-homology

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