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Electroporation: Introduction and Expression of Transgenes in Plant Protoplasts

  • Henry J. Fisk
  • Abhaya M. Dandekar
Part of the Methods in Molecular Biology™ book series (MIMB, volume 286)

Summary

An optimized protocol for the electroporation-based transfection of tobacco protoplasts is described that routinely results in transgene expression frequencies approaching 90%. The overall efficiency of the procedure depends collectively on numerous key parameters, including protoplast viability; DNA concentration, purity, and topology; carrier DNA; and electrical conditions such as ionic strength of the electroporation buffer, electric field strength, pulse duration, and capacitance. Individual methodologies that address each one of these parameters are presented in sufficient detail to enable successful reproduction of this method along with notes that describe helpful tips.

Key Words

Electroporation plant transformation protoplast transfection transient expression 

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Copyright information

© Humana Press Inc. 2005

Authors and Affiliations

  • Henry J. Fisk
    • 1
  • Abhaya M. Dandekar
    • 1
  1. 1.Department of PomologyUniversity of CaliforniaDavis

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