By Jac A. Nickoloff
Harvard college, Boston, Massachusetts. handbook of thought, instrumentation, and step by step protocols within the electroporation of plant cells. For molecular geneticists and different researchers. 24 participants, 22 U.S.
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Extra resources for Plant Cell Electroporation And Electrofusion Protocols (Methods in Molecular Biology)
Sample text
Table 3 shows generators available with increasing flexibility and cost options. The maximum voltage is typically 2500 V. The time constant for fixed pulse length is typically 5 ms. 2. Square- Wave Generators Square-wave pulses appearto have advantagesfor certain applications, such as transfection of mammalian cell lines and plant protoplasts, though no generalization can be made. Each cell line needs to be individually investigated to determine whether use of square-wave pulses would be advantageous.
However, recent advances in solid-state switching technology have lowered costs. A square-wave generator is now available that can deliver up to 3000 V into a 20-n load at costs comparable to exponential discharge units (see Table 1). 3. Generators for Electrofusion If nonelectrical means of cell-cell contact are used, any electroporation generator can also be used for electrofusion. If it is desirable to induce cell-cell contact by dielectrophoresis, the generator needs to produce an alternating wave form (ac) over a longer period of time, typically seconds, before the fusion pulse is applied.
However, if the instrument and chamber stand are designed correctly, this should be of no consequence. It should be noted here that arcing in the chamber occurs mostly at high field strengths (above 10 kV/cm) and is a statistical effect. The resistance R, is a timing resistor that, typically, can be selected to adjust the pulse length. Maintaining a low value, relative to the chamber resistance R,, serves the function of determining pulse length. Often, the size of the capacitance can be changed by connecting one or more capacitors in parallel.