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Oat pp 55–70Cite as

Manipulation of Oat Protoplasts for Transient Expression Assays

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1536))

Abstract

Oat protoplasts are a useful and convenient system to study transient expression using whole cells. Nucleic acid can rapidly be introduced into live cells, and, depending on the assay, results can be collected the same day. Compared to plant tissue, oat cell suspension cultures provide a simple, high yielding, and consistent means to isolate protoplasts. Here, we describe how to generate an oat cell suspension culture from callus grown on solidified medium, and how to maintain the oat cells in cell suspension culture for protoplast preparation. Following the culturing procedure, we describe how to isolate oat protoplasts from cell suspension culture by enzymatic digestion of the cell walls and to transiently express nucleic acid (DNA or RNA) into the cells by electroporation.

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References

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Authors and Affiliations

  1. University of Wisconsin-Madison, Madison, WI, USA

    Robyn Roberts, Jincan Zhang, Nicole Mihelich, Danielle Savino & Aurélie M. Rakotondrafara

Authors
  1. Robyn Roberts
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  2. Jincan Zhang
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  3. Nicole Mihelich
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  4. Danielle Savino
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  5. Aurélie M. Rakotondrafara
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Corresponding author

Correspondence to Aurélie M. Rakotondrafara .

Editor information

Editors and Affiliations

  1. Department of Functional Genomics, Plant Breeding & Acclimatization Inst. National Research Institute, Blonie, Poland

    Sebastian Gasparis

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Roberts, R., Zhang, J., Mihelich, N., Savino, D., Rakotondrafara, A.M. (2017). Manipulation of Oat Protoplasts for Transient Expression Assays. In: Gasparis, S. (eds) Oat. Methods in Molecular Biology, vol 1536. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6682-0_5

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  • DOI: https://doi.org/10.1007/978-1-4939-6682-0_5

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6680-6

  • Online ISBN: 978-1-4939-6682-0

  • eBook Packages: Springer Protocols

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