Electroporation Protocol

Bednarz J, Teifel M, Friedl P, Engelmann K. Immortalization of human corneal endothelial cells using electroporation protocol optimized for human corneal endothelial and human retinal pigment epithelial cells. Acta Ophthalmol Scand 78 130-136 (2000). 

FIGURE 17.2 Enhancement ratios of total amount of estradiol penetrated and skin deposition from ultradeformable liposomes after an electroporation protocol (5 pulses of 100 V, 100 ms, and 1 min spacing) relative to passive diffusion from the vesicles. 

To produce stable and long-lasting knockdown, viral vectors have been developed to efficiently express siRNAs or short hairpin RNAs (shRNAs) in a wide variety of mammalian cells (Fig. 8). Expression vectors include standard plasmids as well as those made from adenoviruses, adeno-associated viruses (AAVs), onco-retroviruses, and lentiviruses. The viral expression systems have the added benefit of being able to deliver siRNA, shRNA, or miRNA expression cassettes efficiently into cell types that are otherwise difficult to transfect via common transfection or electroporation protocols.f ... 

Electroporation Protocols Preclinical and Clinical Gene Medicine, edited by Shulin Li,... 

Dictyostelium discoideum cells can be transformed with plasmid DNA by convenient methods. The first attempts at this, using the calcium phosphate method, were reported in the early 1980s [17, 18]. Protocols were subsequently optimized by Firtel and coworkers [19, 20] and modified in many laboratories (e.g. Ref [21]). Up to 2000 transformants can be recovered from 10 D. discoideum cells [20], with electroporation protocols having been reported [22, 23]. It has also been noted that the choice of transformation method can strongly influence the yield of heterolo-gously expressed proteins [24]. Hence, both calcium co-precipitation and electroporation should be compared to generate optimized expression strains for the production of biopharmaceutical proteins in D. discoideum. 

D PAGE Sample Preparation and Fractionation, Volume 2, edited by Anton Posch, 2008 2D PAGE Sample Preparation and Fractionation, Volume I, edited by Posch, 2008 Electroporation Protocols Preclinical and Clinical Gene Medicine, edited by Shulin Li. 2008 Phylogenomics edited by William J. Murphy, 2008 Affinity Chromatc raphy Methods and Protocols, Second Edition, edited by Michael Zachariou, 2008 Drosophila Methods and Protocols, edited by Christian Dahmann, 2008... 

Turgeon N, Laflamme C, Ho J, Duchaine C. (2006). Elaboration of an electroporation protocol for Bacillus cereus ATCC 14579. Journal of Microbiological Methods 67 543-548. 

Animal Cell Electroporation and Electrofusion Protocols, edited by Jac A. Nickoloff. 1995 47. Electroporation Protocols for Microorganisms, edited by Jac A. Nickoloff 1995... 

Cells are routinely passaged two days prior to electroporating. Cells are ready for electroporating when their density is optimal. Usually one 10-cm plate at approx 80% confluency will provide enough cells for 1-2 electroporations. Our standard electroporation protocol is given below. 

Procedures for transfection by calcium phosphate-DNA coprecipitation and by electroporation are given in our earlier reviews (Cherbas et al. 1994 Cherbas and Cherbas 1998). The electroporation protocol for Kc cells (Cherbas et al. 1994) can be modified for S2 cells by increasing the voltage from 440 V/cm (Kc) to 715 V/cm (S2) (Cherbas and Cherbas 1998) the higher voltage also works well for S3 cells, for the shibire line EH34A3, and for the haploid line D (K. Klueg and M.A.T. Muskavitch, pers. comm.). 

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