African Journal of Biotechnology Vol. 2 (4), pp. 86-87, April 2003

A home made kit for plasmid DNA mini-preparation

Simeon Oloni KOTCHONI^1*, Emma Wanjiru GACHOMO², Eriola BETIKU^3,Y and Olusola Olusoji SHONUKAN⁴

¹Department of Plant Molecular Biology, Institute of Botany, Kirschallee 1, University of Bonn, D-53115 Germany.

²Institute for Plant Diseases, Nussallee 9, University of Bonn, D-53115 Germany.

³Chemical Engineering Department, Faculty of Technology, Obafemi Awolowo University, Ile-Ife, Nigeria.

⁴Department of Microbiology, Faculty of Science, Obafemi Awolowo University, Ile-Ife, Nigeria.

^YCurrent Address: German Research Centre for Biotechnology (GBF), Mascheroder Weg 1, Braunschweig, D-38124 Germany.

* Corresponding author; Tel: +49-228739580, fax: +49-228-732689, e-mail: kotchoni@uni-bonn.de

Accepted 26 March 2003

Many methods have been used to isolate plasmid DNA, but some of them are time consuming especially when extracting a large number of samples. Here, we developed a rapid protocol for plasmid DNA extraction based on the alkaline lysis method of plasmid preparation (extraction at pH 8.0). Using this new method, a good plasmid preparation can be made in approximately one hour. The plasmids are suitable for any subsequent molecular applications in the laboratory. By applying the recommendations to avoid contaminations and to maximize the plasmid yield and quality during extraction, this protocol could be a valuable reference especially when analyzing a large number of samples.

Key words: Plasmid extraction, PCR, restriction enzymes, sequencing, contamination.