By Jac A. Nickoloff
This extraordinary number of well-tested protocols for the electroporation of proteins and DNA into insect, fish, and mammalian cells additionally comprises confirmed animal cellphone electrofusion suggestions for reviews of somatic cellphone genetics and improvement, and for producing monoclonal antibodies. it's exotic via its insurance of significant version mobile forms from many organisms and tissue kinds and its unique directions for the expansion and practise of particular cells to accomplish optimal transfection. Its broad reference lists, citations of different move tools, suggestion on pitfalls to prevent, and outlines of anticipated effects confirm quite simply reproducible effects even for rookies.
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Additional info for Animal Cell Electroporation and Electrofusion Protocols
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. In this chapter, the term electro cell manipulation (ECM) shall describe both techniques.
A (1989) Optimization of electroporatlon transfection of mammalian cells. Anal. Biochem 180,269-275. for 40 Hui 8. , Klenchin, V. , and Chizmadzhev, Y. A. (1992) Electroporation and electrophoretic DNA transfer mto cells The effect of DNA interaction with electropores Bzophys. J. 63,1320-1327 9. Xie, T. , and Tsong, T Y. (1990) Study of mechamsms of electrrc fieldinduced DNA transfectron I DNA entry by surface binding and diffusion through membrane pores. Biophys. J. 58, 13-19. 10 Nlckoloff, J.
A comparison of plant protoplast fusion yields using a large-volume parallel plate chamber made of SS or a gold-plated concentric ring chamber (both for Petri dishes) showed a consistently higher yield for the gold-plated chamber (10). Table 6 shows a survey of commercially available chambers. 6. Measuring ECM Parameters By following an established protocol, it is generally not necessary to measure the ECM parameters, especially if the same types of generator Instrumentation 57 Fig. 9. Petri dish electrodes for large-volume electroporation of mammalian cells in suspension or adherence, and electroporation of fish eggs.