Abstract
Modern DNA assembly techniques are known for their potential to link multiple large DNA fragments together into even larger constructs in single pot reactions that are easier to automate and work more reliably than traditional cloning methods. The simplicity of the chemistry is in contrast to the increased work needed to design optimal reactions that maximize DNA fragment reuse, minimize cost, and organize thousands of potential chemical reactions. Here we examine available DNA assembly methods and describe through example, the construction of a complex but not atypical combinatorial and hierarchical library using protocols that are generated automatically with the assistance of modern synthetic biology software.
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Acknowledgments
Michael Matena and Adam Thomas for development of TeselaGen’s Hierarchical Design Editor; Thomas Rich and Tiffany Dai for development of TeselaGen’s Open Vector Editor; Rodrigo Pavez, Tim Thimmaiah, and Nick Elsbree for TeselaGen software platform development to support DNA design; and Katy Basile and Matthew Gibson for intellectual property consultation and support. Portions of this work were funded by NSF 1430986.
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Fero, M.J., Craft, J.K., Vu, T., Hillson, N.J. (2020). Combinatorial-Hierarchical DNA Library Design Using the TeselaGen DESIGN Module with j5. In: Chandran, S., George, K. (eds) DNA Cloning and Assembly. Methods in Molecular Biology, vol 2205. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0908-8_2
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DOI: https://doi.org/10.1007/978-1-0716-0908-8_2
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