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Transgenic Plants pp 19–35Cite as

The Use of an Automated Platform to Assemble Multigenic Constructs for Plant Transformation

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1864))

Abstract

Compared to traditional means, modern DNA assembly methods allow cloning of large, multigenic vectors for plant transformation in rapid fashion. These methods are often robust and efficient and can assemble multiple DNA fragments into a single vector in one reaction. Here we describe the use of an automated DNA assembly platform for the generation of complex, multigenic T-DNA binary vectors using a hierarchical Golden Gate cloning strategy. These DNA constructs contained diverse DNA elements for the expression of multiple genes for trait stacking in the crop of interest. This platform streamlines the DNA assembly and validation process through high-efficiency cloning methods, integrated automation equipment, and increased throughput. The implementation of this platform removes bottlenecks for routine molecular biology and opens new possibilities for downstream experimental idea testing.

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Acknowledgments

The authors would like to thank Bill Moskal for his intellectual input on the integration of an automated platform, Siobhan Davis for her collaboration on sample management and archiving, Ryan Blue and Patrick Westfall for their intellectual input on DNA assembly methods, and Ann Owens Merlo and Tom Meade for their leadership and support.

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Authors and Affiliations

  1. Corteva Agriscience™, Agriculture Division of DowDuPont™, Indianapolis, IN, USA

    David G. J. Mann, Scott A. Bevan, Anthony J. Harvey & Rachelle A. Leffert-Sorenson

Authors
  1. David G. J. Mann
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  2. Scott A. Bevan
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  3. Anthony J. Harvey
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  4. Rachelle A. Leffert-Sorenson
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Corresponding author

Correspondence to David G. J. Mann .

Editor information

Editors and Affiliations

  1. Corteva Agriscience™, Agriculture Division of DowDuPont™, Johnston, IA, USA

    Sandeep Kumar

  2. Corteva Agriscience™, Agriculture Division of DowDuPont™, Johnston, IA, USA

    Pierluigi Barone

  3. Corteva Agriscience™, Agriculture Division of DowDuPont™, Johnston, IA, USA

    Michelle Smith

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Mann, D.G.J., Bevan, S.A., Harvey, A.J., Leffert-Sorenson, R.A. (2019). The Use of an Automated Platform to Assemble Multigenic Constructs for Plant Transformation. In: Kumar, S., Barone, P., Smith, M. (eds) Transgenic Plants. Methods in Molecular Biology, vol 1864. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8778-8_2

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  • DOI: https://doi.org/10.1007/978-1-4939-8778-8_2

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8777-1

  • Online ISBN: 978-1-4939-8778-8

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