ISSN 2736-1756
Advanced Journal of Microbiology Research Vol. 2003
Available online at http://internationalscholarsjournals.org/journal/ajmr
© 2003 International Scholars Journals
A home made kit for plasmid DNA mini-preparation
Simeon Oloni KOTCHONI1 , Emma Wanjiru GACHOMO2, Eriola BETIKU3, 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.
Accepted 26 March 2003
Abstract
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.