Electroporation of DNA and RNA into Plant Protoplasts

Michael Fromm, Judy Callis, Loverine P. Taylor, Virginia Walbot

Research output: Contribution to journalArticle

84 Citations (Scopus)

Abstract

This chapter describes the procedures for electrically transferring DNA and RNA into monocot and dicot plant protoplasts. The chapter presents electrical fundamentals and equipment design and the detailed protocols and parameters that affect electroporation-mediated gene transfer. Several of the features described in the chapter for plant cells are relevant to the electroporation of animal cells. Capacitors are electrical devices for storing electrical charge. Capacitors differ from batteries in that capacitors store charge through the use of electric fields, while batteries store charge through the use of chemical reactions. The three important parameters of a capacitor are as follows: (1) the capacitance, typically in microfarads (μF), (2) the maximum safe working voltage of the capacitor, and (3) the polarity of the capacitor (many nonelectrolytic capacitors do not have a polarity). A case surrounding the electrical components of the electroporation device is essential to prevent accidental contact with any of the components such as resistors, switches, oscilloscope attachment, and others. The chapter describes several steps to solve electroporation/expression problems to gain familiarity with electroporation.

Original languageEnglish (US)
Pages (from-to)351-366
Number of pages16
JournalMethods in enzymology
Volume153
Issue numberC
DOIs
StatePublished - Jan 1987

Fingerprint

Plant RNA
Electroporation
Protoplasts
Capacitors
RNA
DNA
Equipment Design
Equipment and Supplies
Plant Cells
Gene transfer
Resistors
Chemical reactions
Animals
Capacitance
Cells
Switches
Electric fields
Genes
Electric potential

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Fromm, M., Callis, J., Taylor, L. P., & Walbot, V. (1987). Electroporation of DNA and RNA into Plant Protoplasts. Methods in enzymology, 153(C), 351-366. https://doi.org/10.1016/0076-6879(87)53064-6

Electroporation of DNA and RNA into Plant Protoplasts. / Fromm, Michael; Callis, Judy; Taylor, Loverine P.; Walbot, Virginia.

In: Methods in enzymology, Vol. 153, No. C, 01.1987, p. 351-366.

Research output: Contribution to journalArticle

Fromm, M, Callis, J, Taylor, LP & Walbot, V 1987, 'Electroporation of DNA and RNA into Plant Protoplasts', Methods in enzymology, vol. 153, no. C, pp. 351-366. https://doi.org/10.1016/0076-6879(87)53064-6
Fromm, Michael ; Callis, Judy ; Taylor, Loverine P. ; Walbot, Virginia. / Electroporation of DNA and RNA into Plant Protoplasts. In: Methods in enzymology. 1987 ; Vol. 153, No. C. pp. 351-366.
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