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Applied and Environmental Microbiology, January 2009, p. 414-418, Vol. 75, No. 2
0099-2240/09/$08.00+0     doi:10.1128/AEM.01358-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Optimized Method for Preparation of DNA from Pathogenic and Environmental Mycobacteria{triangledown}

Michael Käser,1* Marie-Thérèse Ruf,1 Julia Hauser,1 Laurent Marsollier,2 and Gerd Pluschke1

Swiss Tropical Institute, Socinstrasse 57, 4002 Basel, Switzerland,1 Université d'Angers, Centre Hospitalier Universitaire, 4 Rue Larrey, 49033 Angers Cedex 1, France2

Received 17 June 2008/ Accepted 11 November 2008

Genomic studies on pathogenic and environmental mycobacteria are of growing interest for understanding of their evolution, distribution, adaptation, and host-pathogen interaction. Since most mycobacteria are slow growers, material from in vitro cultures is usually scarce. The robust mycobacterial cell wall hinders both experimental cell lysis and efficient DNA extraction. Here, we compare elements of several DNA preparation protocols and describe a method that is economical and practical and reliably yields large amounts—usually 10-fold increased compared to earlier protocols—of highly pure genomic DNA for sophisticated downstream applications. This method was optimized for cultures of a variety of pathogenic and environmental mycobacterial species and proven to be suitable for direct mycobacterial DNA extraction from infected insect specimens.

* Corresponding author. Mailing address: Swiss Tropical Institute, Molecular Immunology, Socinstr. 57, 4002 Basel, Switzerland. Phone: 41-61-2848-171. Fax: 41-61-2848-101. E-mail: m.kaeser{at}unibas.ch

{triangledown} Published ahead of print on 1 December 2008.

Applied and Environmental Microbiology, January 2009, p. 414-418, Vol. 75, No. 2
0099-2240/09/$08.00+0     doi:10.1128/AEM.01358-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.



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