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From 2011.igem.org

• Home
• About us
  • The Team
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  • Gallery
• The Project
  • Abstract
  • Introduction
  • The Plug 'n' Play
    • Assembly system
    • Customization
  • Other assembly systems
  • Achievements
  • Attributions
• Results
  • Background
  • Characterization
  • Proof of concept
    • Mammalian Cells
    • Fungi
  • Data page
  • Submitted Biobricks
  • Collaboration
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• Notebook
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  • Protocols
• Our Partners
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  • Acknowledgements

	1 Gateway system 2 Gibson Assembly 3 In fusion 4 Sandwich 5 3A	Gibson Assembly Gibson Assembly is an isothermal, single-reaction method for assembling multiple overlapping DNA molecules, which was developed by Daniel G. Gibson at the J. Craig Venter Institute in 2009. The assembly system employs 5´-T5 exonuclease, PhusionDNA polymerase, and Taq lig and can be used to assemble both ssDNA and dsDNA. This methode makes it possible to join DNA molecules there are as large as 583kb and clone joined products in ''E. coli'' up to 300kb. The way it's done The overlapping DNA molecules We describe an isothermal, single-reaction method for assembling multiple overlapping DNA molecules by the concerted action of a 5¢ exonuclease, a DNA polymerase and a DNA ligase. First we recessed DNA fragments, yielding single-stranded DNA overhangs that specifically annealed, and then covalently joined them. This assembly method can be used to seamlessly construct synthetic and natural genes, genetic pathways and entire genomes, and could be a useful molecular engineering tool.