Animal Cell Electroporation and Electrofusion Protocols by Jac A. Nickoloff

By Jac A. Nickoloff

This fantastic number of well-tested protocols for the electroporation of proteins and DNA into insect, fish, and mammalian cells additionally comprises confirmed animal mobile electrofusion recommendations for reviews of somatic telephone genetics and improvement, and for producing monoclonal antibodies. it truly is distinctive by means of its insurance of significant version mobile forms from many organisms and tissue forms and its certain directions for the expansion and education of particular cells to accomplish optimal transfection. Its large reference lists, citations of other move tools, recommendation on pitfalls to prevent, and outlines of anticipated effects make sure with no trouble reproducible effects even for novices.

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As a consequence, cell viability imposes an upper limit on the transfection efficiency. 3. Experimental Evidence Several experiments using fluorescent molecules as tracers have shown that macromolecule leakage or intake during electroporation is indeed proportional to the quantity (E - Eb)T (4,12,13). Figure 1 shows the uptake of fluorescein isothiocyanate-labeled dextran by mouse C3W 10T,,Z cells as a function of ET. Because cells take up dextran spontaneously, the x-axis intercept does not correspond to the value of EbT.

And Teissre, J. (1994) Control by pulse parameters of electric field-mediated gene transfer in mammalian cells. Biophys. J. 66,524-531. 7 Andreason, G. L. and Evans, G. 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.

And Tsong, T. Y. (1990) Study of mechanisms of electric field-mduced DNA transfection. II. Transfection by low amplitude low frequency alternating electric fields. Biophys. J. 58,897-903. 19. Chang, D. , Gao, P. , 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.

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