By Jac A. Nickoloff
This brilliant selection of well-tested protocols for the electroporation of proteins and DNA into insect, fish, and mammalian cells additionally comprises confirmed animal mobilephone electrofusion suggestions for stories of somatic mobile genetics and improvement, and for producing monoclonal antibodies. it truly is uncommon via its insurance of significant version phone varieties from many organisms and tissue forms and its unique directions for the expansion and training of particular cells to accomplish optimal transfection. Its vast reference lists, citations of different move tools, suggestion on pitfalls to prevent, and outlines of anticipated effects determine comfortably reproducible effects even for newbies.
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Extra resources for Animal Cell Electroporation and Electrofusion Protocols
14. Xie, T. D. and Tsong, T Y. (1992) Study of mechanisms of electric field-induced DNA transfection III Electrrc parameters and other conditions for effective transfectlon. Biophys. J. 63,28-34. 15. Kubimec, R. , and Hui, S. W. (1990) Effects of pulse length and pulse strength on transfectlon by electroporatlon. BioTechniques 8, l-3. 16. Chang, D. , Chassy, B. , Saunders, J. , and Sowers, A. E. (1992) Gwdefor Electroporation and Electrojksion Academic, San Diego. 17. Baker, P. F. and Knight, D.
Generators with Other 5. Chambers There are many choices in chambers for ECM. In general, chambers need to create the required field strength from the voltage delivered to the electrodes by the generator; they need to contain the appropriate volume, and need to be sterilized or sterilizable, easily filled, emptied, and if reused, easily cleaned. Table 5 gives the main trade-off parameters in the selection of chambers. The following describes only the more frequently used chambers in the field. Instrumentation 51 Table 5 Chamber Trade-off Parameters Small volume Disposable Homogeneous field Visualization of cells Batch process Aluminum electrodes Presterilized Small gap Fig.
Notes 1. Electroporation can be used to mtroduce a variety of proteins mto cells, including monoclonal antibodies (MAb) (22,23), functional enzymes (24), other endonucleases in addition to restriction enzymes (25), and a host of other proteins and protein products (26-28). 2. Both the Bio-Rad Gene Pulser and the BRL Cell Porator are effective in the introduction of enzymes into the cell, but the most effective conditions for the BRL machine also result in relatively high levels of cell death (10).