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An Automated Protoplast Transformation System

  • Scott C. LenaghanEmail author
  • C. Neal StewartJr.Email author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1917)

Abstract

Efficient plant protoplast production from cell suspension cultures, leaf, and stem tissue allows for single-cell plant biology. Since protoplasts do not have cell walls, they can be readily transformed to enable rapid assessment of regulatory elements, synthetic constructs, gene expression, and more recently genome-editing tools and approaches. Historically, enzymatic cell wall digestion has been both expensive and laborious. Protoplast production, transformation, and analysis of fluorescence have recently been automated using an integrated robotic system. Here we describe its use for bulk protoplast isolation, counting, transformation, and analysis at very low cost for high-throughput experiments.

Key words

Tobacco Protoplasts Transformation Enzymatic digestion High-throughput screening Automation Robotics 

Notes

Acknowledgement

This research was supported by Advanced Research Projects Agency-Energy (ARPA-E) Award No. DE-AR0000313, with additional funding and assistance from ThermoFisher.

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Copyright information

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

Authors and Affiliations

  1. 1.Department of Food ScienceUniversity of TennesseeKnoxvilleUSA
  2. 2.Department of Mechanical, Aerospace, and Biomedical EngineeringUniversity of TennesseeKnoxvilleUSA
  3. 3.Department of Plant SciencesUniversity of TennesseeKnoxvilleUSA

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