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High efficiency transformation of E.coli by high voltage electroporation
Open accessAbstract
E. coli can be transformed to extremely high efficiencies by subjecting a mixture of cells and DNA to brief but intense electrical fields of exponential decay waveform (electroporation). We have obtained 10(9) to 10(10) transformants/micrograms with strains LE392 and DH5 alpha, and plasmids pUC18 and pBR329. The process is highly dependent on two characteristics of the electrical pulse: the electric field strength and the pulse length (RC time constant). The frequency of transformation is a linear function of the DNA concentration over at least six orders of magnitude; and the efficiency of transformation is a function of the cell concentration. Most of the surviving cells are competent with up to 80% transformed at high DNA concentration. The mechanism does not appear to include binding of the DNA to the cells prior to entry. Possible mechanisms are discussed and a simple procedure for the practical use of this technique is presented.
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Cited by
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Oettinger, M. A., Schatz, D. G., Gorka, C., & Baltimore, D. (1990) · Science · doi:10.1126/science.2360047 · Cited by 1,456
Hoogenboom, H. R., Griffiths, A. D., Johnson, K. S., Chiswell, D. J., Hudson, P., & Winter, G. (1991) · Nucleic Acids Research · doi:10.1093/nar/19.15.4133 · Cited by 1,020
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Packer, M. S., & Liu, D. R. (2015) · Nature Reviews Genetics · doi:10.1038/nrg3927 · Cited by 991
The Agent of Bacillary Angiomatosis: An Approach to the Identification of Uncultured Pathogens
Relman, D. A., Loutit, J. S., Schmidt, T. M., Falkow, S., & Tompkins, L. S. (1990) · New England Journal of Medicine · doi:10.1056/nejm199012063232301 · Cited by 901
[12] High-efficiency transformation of yeast by electroporation
(1991) · Methods in Enzymology · doi:10.1016/0076-6879(91)94015-5 · Cited by 788
Metadata
- Authors
- Dower, W. J., Miller, J. F., & Ragsdale, C. W.
- Venue
- Nucleic Acids Research
- Publisher
- Oxford University Press (OUP)
- Published
- 1988-07-11
- Volume
- 16
- Issue
- 13
- Pages
- 6127-6145
- ISSN
- 0305-1048 (ISSN-L)
- 1362-4962
- OpenAlex
- W2152845055
- Cited by
- 2,809
- References
- 0
- FWCI
- 77.92
- Type
- journal article
- Language
- en
- Open access
- green
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- Dower, W. J., Miller, J. F., & Ragsdale, C. W. (1988). High efficiency transformation of E.coli by high voltage electroporation. Nucleic Acids Research. https://doi.org/10.1093/nar/16.13.6127
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