Abstract
Gene synthesis is a fundamental technology underpinning much research in the life sciences. In particular, synthetic biology and biotechnology utilize gene synthesis to assemble any desired DNA sequence, which can then be incorporated into novel parts and pathways. Here, we describe SpeedyGenes, a gene synthesis method that can assemble DNA sequences with greater fidelity (fewer errors) than existing methods, but that can also be used to encode extensive, statistically designed sequence variation at any position in the sequence to create diverse (but accurate) variant libraries. We summarize the integrated use of GeneGenie to design DNA and oligonucleotide sequences, followed by the procedure for assembling these accurately and efficiently using SpeedyGenes.
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References
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Acknowledgements
We thank the Biotechnology and Biological Sciences Research Council for financial support (grant BB/M017702/1); Prof Nick Turner, Dr. Ian Rowles, and Dr. Timothy Eyes for useful discussions; and Mrs. Hannah Currin for preparation of figures. This is a contribution from the Manchester Centre for Synthetic Biology of Fine and Speciality Chemicals (SYNBIOCHEM).
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Currin, A., Swainston, N., Day, P.J., Kell, D.B. (2017). SpeedyGenes: Exploiting an Improved Gene Synthesis Method for the Efficient Production of Synthetic Protein Libraries for Directed Evolution. In: Hughes, R. (eds) Synthetic DNA. Methods in Molecular Biology, vol 1472. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6343-0_5
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DOI: https://doi.org/10.1007/978-1-4939-6343-0_5
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