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.
ASJC Scopus subject areas
- Molecular Biology