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Nucleic Acids Research, 2000, Vol. 28, No. 8 E37-00
© 2000 Oxford University Press

Amplifiable DNA from Gram-negative and Gram-positive bacteria by a low strength pulsed electric field method

Frank Vitzthum, Georg Geiger, Hans Bisswanger1, Bentsian Elkine, Herwig Brunner and Jürgen Bernhagen*

Laboratory of Biochemistry, Institute for Interfacial Engineering, University of Stuttgart and Fraunhofer-IGB, Nobelstraße 12, D-70569 Stuttgart, Germany and 1Physiological-chemical Institute, University of Tübingen, Hoppe-Seyler-Straße 4, D-72076 Tübingen, Germany

An efficient electric field-based procedure for cell disruption and DNA isolation is described. Isoosmotic suspensions of Gram-negative and Gram-positive bacteria were treated with pulsed electric fields of <60 V/cm. Pulses had an exponential decay waveform with a time constant of 3.4 µs. DNA yield was linearly dependent on time or pulse number, with several thousand pulses needed. Electrochemical side-effects and electrophoresis were minimal. The lysates contained non-fragmented DNA which was readily amplifiable by PCR. As the method was not limited to samples of high specific resistance, it should be applicable to physiological fluids and be useful for genomic and DNA diagnostic applications.

* To whom correspondence should be addressed. Tel: +49 711 970 4020; Fax: +49 711 970 4300; Email: jbe@igb.fhg.de


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