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Bricks and blueprints: methods and standards for DNA assembly

Abstract

DNA assembly is a key part of constructing gene expression systems and even whole chromosomes. In the past decade, a plethora of powerful new DNA assembly methods — including Gibson Assembly, Golden Gate and ligase cycling reaction (LCR) — have been developed. In this Innovation article, we discuss these methods as well as standards such as the modular cloning (MoClo) system, GoldenBraid, modular overlap-directed assembly with linkers (MODAL) and PaperClip, which have been developed to facilitate a streamlined assembly workflow, to aid the exchange of material between research groups and to create modular reusable DNA parts.

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Figure 1: Examples of type IIS restriction endonuclease-based methods and standards.
Figure 2: Site-specific recombination methods and standards.
Figure 3: Examples of long-overlap-based assembly methods and standards.

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Acknowledgements

The authors thank M. Jamilly for contributing to initial discussions and apologize to those whose work was not discussed owing to space limitations. Research on DNA assembly in the groups of T.E. and G.S.B. is supported by the UK Engineering and Physical Research Council (EPSRC) grant EP/J02175X/1 and EU FP7 grant KBBE.2011.5-289326. M.S. is supported by Marie Curie Intra-European Fellowship 628019.

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Correspondence to Geoffrey S. Baldwin or Tom Ellis.

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Casini, A., Storch, M., Baldwin, G. et al. Bricks and blueprints: methods and standards for DNA assembly. Nat Rev Mol Cell Biol 16, 568–576 (2015). https://doi.org/10.1038/nrm4014

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