High efficiency transformation of E.coli by high voltage electroporation

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ID: 290206
1988
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Abstract
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.
Reference Key
openalex_W2152845055 Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors William J. Dower, Jeff F. Miller, Clifton W. Ragsdale
Journal Nucleic Acids Research
Year 1988
DOI
10.1093/nar/16.13.6127
URL
https://doi.org/10.1093/nar/16.13.6127
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