By Jac A. Nickoloff
This brilliant choice of well-tested protocols for the electroporation of proteins and DNA into insect, fish, and mammalian cells additionally comprises confirmed animal phone electrofusion strategies for reports of somatic phone genetics and improvement, and for producing monoclonal antibodies. it truly is individual through its insurance of significant version telephone varieties from many organisms and tissue kinds and its certain directions for the expansion and practise of particular cells to accomplish optimal transfection. Its broad reference lists, citations of other move tools, recommendation on pitfalls to prevent, and outlines of anticipated effects be sure without problems reproducible effects even for newcomers.
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Additional info for Animal Cell Electroporation and Electrofusion Protocols
And Maxwell, B. L (1991) High efficiency gene transfection by electroporation using a radio-frequency electric field. Biochim. Btophys. Actu 1992,153-160. 20 Tekle, E , Austumlan, R. , and Chock, P. B (1991) Electroporatron by using bipolar oscillating electric field. an improved method for DNA transfectlon of NIH 3T3 cells. Proc. Natl. Acad. Sci USA 88,4230-4234 CHAPTER3 Instrumentation Gunter A. Hofmann 1. Introduction The techniques of electroporation and electrofusion require that cells be subjected to brief pulses of electric fields of the appropriate amplitude, duration, and wave form.
They are also available in different gap sizes. 52 Hofmann Fig. 4. Reusable parallel plate electrode assembly to fit into spectrophotometer cuvets. Note that chemical cleaning or even autoclaving might not remove cell debris, transformant, and medium breakdown products that might have been deposited onto the electrodes during a pulse. Only a good mechanical cleaning will remove the debris. Electrodes on microslides are used to visualize the fusion process under a microscope. Parallel wires (Fig.
Yates, B. , and Morgan, W F. (1993) Analysis of restriction enzyme-induced chromosomal aberrations by fluorescence in situ hybridization Environ. Mol. Mutagen. 22,26-33. 2 Andreason, G. L. and Evans, G. A. (1988) Introductton and expression of DNA molecules in eukaryotic cells by electroporation (published erratum appears in BioTechmques 6,853) BioTechnrques 6,650-660. 3. , and Teissie, J. (1992) Fast kinetics studies of Escherichia coli electrotransformation. Eur. J Biochem. 209,431-436. 4. Morgan, W.
Animal Cell Electroporation and Electrofusion Protocols by Jac A. Nickoloff