Animal Cell Electroporation and Electrofusion Protocols by Jac A. Nickoloff

By Jac A. Nickoloff

This very good selection of well-tested protocols for the electroporation of proteins and DNA into insect, fish, and mammalian cells additionally comprises confirmed animal telephone electrofusion recommendations for reviews of somatic mobilephone genetics and improvement, and for producing monoclonal antibodies. it truly is distinctive by means of its assurance of significant version mobile kinds from many organisms and tissue varieties and its specific 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 verify comfortably reproducible effects even for newcomers.

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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. E. (1983) High-voltage techniques for gaming access to the interior of cells application to the study of exocytosis and membrane turnover. Methods Enzymol. 98,28-37. 18. Xie, T. D. and Tsong, T. Y. (1990) Study of mechanisms of electric field-mduced DNA transfection. II. Transfection by low amplitude low frequency alternating electric fields.

6 40 130 Partial No Yes Operator shock proof 120 Partial No Yes Safety interlock 40 Partial Yes No Safety interlock No 1 yr No 1 Yr No 1 Yr BTX ECM@ 600 1 >lOOO Full Yes Yes Operator shock, arc, and short circmt proof Yes 2 yr tance, is generally desirable. Sometimes the chamber resistance is too low for the timing resistors to be effective. In this case, the chamber resistance itself will determine the pulse length, which then can be adjusted only by varying the capacitance. For the characterization of the pulse into the chamber, only two parameters need to be known: the peak voltage and the l/e pulse length.

Disposable electroporation Large volume Reusable Inhomogeneous field Cells obscured Flowthrough Stainless steel or noble material Sterilizable Large gap cuvet with molded-in aluminum electrodes. 1. Small-Volume Chambers Disposable, presterilized cuvets with molded-in aluminum electrodes (Fig. 3) are most frequently used for electroporation. They are available with different gap sizes, typically 1 mm (for bacteria), and 2 and 4 mm (for mammalian cells and plant protoplasts). The electric field in these cuvets is quite homogeneous.

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