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Nucleic Acids Res. 1988 Jul 11;16(13):6127-45.

High efficiency transformation of E. coli by high voltage electroporation.

Author information

1
Molecular Biology Group, Bio-Rad Laboratories, Richmond, CA 94804.

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.

PMID:
3041370
PMCID:
PMC336852
DOI:
10.1093/nar/16.13.6127
[Indexed for MEDLINE]
Free PMC Article

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