title: Optimized method for preparation of DNA from pathogenic and environmental mycobacteria.
abstract: 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.
authors.affiliation: Swiss Tropical Institute, Molecular Immunology, Socinstr. 57, 4002 Basel, Switzerland. m.kaeser@unibas.ch
authors.forename: Michael
authors.initials: M
authors.lastname: Käser
authors.forename: Marie-Thérèse
authors.initials: MT
authors.lastname: Ruf
authors.forename: Julia
authors.initials: J
authors.lastname: Hauser
authors.forename: Laurent
authors.initials: L
authors.lastname: Marsollier
authors.forename: Gerd
authors.initials: G
authors.lastname: Pluschke
mesh_terms: D000818:Animals; D004269:DNA, Bacterial / Q000302:isolation & purification*; D007313:Insecta / Q000382:microbiology; D008967:Molecular Biology / Q000379:methods*; D009161:Mycobacterium / Q000235:genetics*
chemical_list: D004269:DNA, Bacterial