The GeneArt® Advanced Genetic Assembly System is a highly efficient kit for the simultaneous and seamless assembly of up to 10 DNA fragments (total length up to 110 Kbp) are assembled into any vector. The system relies on the yeast's ability to collect DNA fragments and recombine them with high efficiency. This greatly reduces in vitro processing of DNA, eliminating the need for enzymatic treatments (such as restriction enzyme digestion and ligation), while allowing precise fusion of DNA sequences. The kit contains materials for transformation and purification from yeast (sterile growth medium) and competent E. coli for plasmid amplification for correct cloning.

•Simple and effective — Clone up to 10 DNA fragments (total length up to 110 Kbp, containing sequences of your choice) simultaneously in a single vector; no restriction enzyme digestion, Joining or Recombination Sites
• Precise and Efficient — Designed to enable you to clone the fragments you need at the site you want, in the direction you want, with up to 90% cloning accuracy, and without Extra sequence will remain
• Flexibility — Use our linear vectors, a vector of your choice, or clone existing DNA fragments with no end homologies without further modification
• Free Tools — Design DNA oligos using our free web interface Nucleotides and more, this interface walks you step-by-step through your entire project
• Multiple Applications — Simplify a variety of synthetic biology and molecular biology by quickly combining, adding, removing, or swapping DNA fragments technology

To clone 1 to 4 DNA fragments of limited size, and if you prefer an in vitro approach, consider using GeneArt® Seamless Cloning and Assembly Kit (Cat. No. A13288).

Easily create new specific constructs from multiple DNA fragments
GeneArt® Advanced Genetic Assembly System utilizes yeast (Saccharomyces cerevisiae) Transformation-coupled recombination (TAR) links existing DNA fragments or chemically synthesized oligonucleotides to a single recombinant molecule. Join DNA fragments and linearized vectors based on shared end homology. If such terminal homology does not exist between the two fragments, it can be "spliced" together with recombinant linkers, synthetic DNA oligonucleotides that provide terminal homology between two unrelated DNA fragments. The process is efficient and seamless, with no additional sequence after assembly. This product has been optimized to clone up to 110 Kb in vectors, although it has been shown to support up to 50 DNA fragments up to 0.5 Mb. Up to 10 DNA fragments.

Simple clone verification
To minimize manipulation of the yeast, recombinant yeast clones use a simplified 10-minute DNA extraction protocol. The extraction step yields an assembly of sufficient molecular weight for colony PCR verification of junctions and direct transformation of E. coli cells for downstream analysis. Proprietary lysis buffer and glass required for extraction are included in the kit. Microbeads. To select recombinant yeast clones, we offer a selective yeast culture kit (Mav203 Yeast Cell CSM Medium; Cat. No. A13292)

Considerations in Choosing Cloning Vectors
The GeneArt® Advanced Genetic Assembly System requires high capacity and. Cloning vectors compatible with yeast and E. coli (i.e., BAC-YAC shuttle vectors) are not included in this product, but we provide one separately called GeneArt® pYES1L Vector with Sapphire Technology™ (Cat. No. A13287) is a ready-to-use linear cloning vector. You can also use your own vector, but it must be compatible with higher-order genetic assembly systems. This compatibility can be achieved through our GeneArt. ® High-level vector transformation cassette (Cat. No. A13291) makes it easy

Cloning efficiency
In GeneArt® high-level assembly, the main factor affecting cloning efficiency is. The DNA element size, the amount of DNA without end homology, the overall final molecular size, and the ends of the PCR fragments (A overhangs or blunt ends) do not affect cloning efficiency. Typical cloning efficiencies for number of fragments with terminal homology assembled into GeneArt® pYES1L Vector with Sapphire Technology™ are as follows:
• For 5 DNA fragments (10 Kb each), >90%
• For 10 DNA fragments (5 Kb each), >90%
• For 10 DNA fragments (10 Kb each), >50%

by "splicing" the DNA oligonucleotides will not have ends Common cloning efficiencies for assembling existing fragments of homology into GeneArt® pYES1L Vector with Sapphire Technology™ are as follows:
• For 1 fragment (10 Kb), >90%
• For 2 DNA fragments ( 10 Kb each), >75%

Computer simulationClone design
GeneArt® A critical step in high-order genetic assembly is correct design Fragments and oligonucleotides with appropriate homology and spacing to ensure successful clone assembly. To simplify and speed up the design process, we provide free online design tools to help you through Computer SimulationDesign Experiments. The tool checks experimental design for compatibility with product specifications, designs DNA oligonucleotides for PCR amplification or splicing of different cloning elements, and shows the user a diagram of the vector and GenBank format that is compatible with Vector NTI® software. Annotated sequences are available for download.

Applications
The GeneArt® Advanced Genetic Assembly System is designed to power cloning and DNA assembly experiments in a variety of molecular biology and synthetic biology applications, as well as other applications. The product can be used to create modular expression vectors with interchangeable parts and can be used to perform a variety of tasks that might otherwise involve multiple steps. Use this kit to easily: construct fusion proteins; delete, replace, or add DNA elements (such as restriction sites); clone larger existing DNA fragments without end homology; and perform many other tasks that require processing of gene sequences. technology.