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Antha-guided Automation of Darwin Assembly for the Construction of Bespoke Gene Libraries

P. Handal Marquez, M. Koch, D. Kestemont, S. Arangundy-Franklin, View ORCID ProfileV. B. Pinheiro
doi: https://doi.org/10.1101/2019.12.17.879486
P. Handal Marquez
1Rega Institute, KU Leuven, Herestraat, 49 box 1041, Leuven, 3000 Belgium
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M. Koch
2Synthace Ltd., The Westworks, 195 Wood Lane, London, W12 7FQ, UK
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D. Kestemont
1Rega Institute, KU Leuven, Herestraat, 49 box 1041, Leuven, 3000 Belgium
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S. Arangundy-Franklin
3Medical Research Council Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK
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V. B. Pinheiro
1Rega Institute, KU Leuven, Herestraat, 49 box 1041, Leuven, 3000 Belgium
4Institute of Structural and Molecular Biology, University College London, Gower Street, London, WC1E 6BT, UK
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  • ORCID record for V. B. Pinheiro
  • For correspondence: v.pinheiro@kuleuven.be
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Abstract

Protein engineering through directed evolution facilitates the screening and characterization of protein libraries. Efficient and effective methods for multiple site-saturation mutagenesis, such as Darwin Assembly, can accelerate the sampling of relevant sequence space and the identification of variants with desired functionalities. Here, we present the automation of the Darwin Assembly method, using a Gilson PIPETMAX™ liquid handling platform under the control of the Antha software platform, which resulted in the accelerated construction of complex, multiplexed gene libraries with minimal hands-on time and error-free, while maintaining flexibility over experimental parameters through a graphical user interface rather than requiring user-driven library-dependent programming of the liquid handling platform. We also present an approach for barcoding libraries that overcomes amplicon length limitations in next generation sequencing and enables fast reconstruction of library reads.

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Posted December 18, 2019.
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Antha-guided Automation of Darwin Assembly for the Construction of Bespoke Gene Libraries
P. Handal Marquez, M. Koch, D. Kestemont, S. Arangundy-Franklin, V. B. Pinheiro
bioRxiv 2019.12.17.879486; doi: https://doi.org/10.1101/2019.12.17.879486
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Antha-guided Automation of Darwin Assembly for the Construction of Bespoke Gene Libraries
P. Handal Marquez, M. Koch, D. Kestemont, S. Arangundy-Franklin, V. B. Pinheiro
bioRxiv 2019.12.17.879486; doi: https://doi.org/10.1101/2019.12.17.879486

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