Abstract
Recovery and purification of ligated plasmid DNA is a critical and limiting step in modern molecular cloning techniques prior to electroporation in E. coli. Four different DNA purification and desalting methods were compared in regard to simple handling, reproducibility, efficiency and cost-effectiveness using picogram and nanogram amounts of DNA. Microcolumn purification of DNA was up to two orders of magnitude more efficient compared to gel filtration, ethanol precipitation or drop dialysis.
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WJ Dower JF Miller CW Ragsdale (1988) ArticleTitleHigh efficiency transformation of E. coli by high voltage electroporation Nucleic Acid Res 16 6127–6145 Occurrence Handle3041370
JF Miller WJ Dower CW Ragsdale (1988) ArticleTitleHigh voltage electroporation of bacteria: genetic transformation of Campylobacter jejuni with plasmid DNA Proc. Natl. Acad. Sci. USA 85 856–860 Occurrence Handle3277182
J Sambrook EF Fritsch T Maniatis (1989) Molecular Cloning: A Laboratory Manual EditionNumber2 Cold Spring Harbor Laboratory Press Cold Spring Harbor, NY
TJ Silhavy ML Berman LW Enquist (1984) Experiments with Gene Fusion Cold Spring Harbor Laboratory Press Cold Spring Harbor, NY
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Revisions requested 14 April 2005; Revisions received 10 May 2005
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Schlaak, C., Hoffmann, P., May, K. et al. Desalting minimal amounts of DNA for electroporation in E. coli: a comparison of different physical methods. Biotechnol Lett 27, 1003–1005 (2005). https://doi.org/10.1007/s10529-005-7867-z
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DOI: https://doi.org/10.1007/s10529-005-7867-z