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

  • David G. J. MannEmail author
  • Scott A. Bevan
  • Anthony J. Harvey
  • Rachelle A. Leffert-Sorenson
Part of the Methods in Molecular Biology book series (MIMB, volume 1864)


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.

Key words

DNA assembly Golden Gate cloning Plasmid T-DNA binary vectors Multigenic construct Plant biotechnology Synthetic biology Automation Liquid handler 



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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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • David G. J. Mann
    • 1
    Email author
  • Scott A. Bevan
    • 1
  • Anthony J. Harvey
    • 1
  • Rachelle A. Leffert-Sorenson
    • 1
  1. 1.Corteva Agriscience™Agriculture Division of DowDuPont™IndianapolisUSA

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