This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Torres, R. D. Articles by Zlotnik, H. Search for Related Content PubMed PubMed Citation Articles by Torres, R. D. Articles by Zlotnik, H.

 Previous Article  |  Next Article 

Clinical and Diagnostic Laboratory Immunology, 09 1996, 601-604, Vol 3, No. 5
Copyright © 1996 by the American Society for Microbiology. All rights reserved.

A rapid and gentle method for isolation of genomic DNA from pathogenic Nocardia spp

RD Torres, CA Oletta and H Zlotnik
Department of Microbiology and Medical Zoology, University of Puerto Rico School of Medicine, San Juan 00936-5067.

The lack of simple and efficient methods for extraction of DNA from Nocardia spp. has hampered molecular manipulation of the DNA for diagnostic purposes. In the present study, a method for the rapid extraction of undegraded genomic nocardial DNA was established. Briefly, 14 pathogenic Nocardia strains were grown at 37 degrees C for 3 to 5 days in Sauton broth containing 0.05% Tween 80. Subsequently, the cultures were treated for 48 h with 1.2 mg of cycloserine per ml (final concentration). Cells were then harvested by centrifugation and treated with a lysis solution containing 3 mg of lysozyme per ml. This was followed by the addition of proteinase K and sodium dodecyl sulfate to final concentrations of 0.2 mg/ml and 0.5%, respectively, and incubation for 1 h at 50 degrees C. DNA was precipitated with isopropanol after phenol-chloroform-isoamyl alcohol extractions and RNase treated before being quantitated and analyzed by agarose gel electrophoresis. The average undegraded DNA yields obtained were 101 micrograms for Nocardia brasiliensis and 121 micrograms for N. asteroides. This DNA was suitable for restriction endonuclease digestion and PCR amplification, which are methods being applied to the characterization and diagnosis of slowly growing organisms such as Nocardia spp.

This article has been cited by other articles:

• Yamamura, H., Hayakawa, M., Nakagawa, Y., Tamura, T., Kohno, T., Komatsu, F., Iimura, Y. (2005). Nocardia takedensis sp. nov., isolated from moat sediment and scumming activated sludge. Int. J. Syst. Evol. Microbiol. 55: 433-436 [Abstract] [Full Text]  
• Yamamura, H., Hayakawa, M., Nakagawa, Y., Iimura, Y. (2004). Characterization of Nocardia asteroides Isolates from Different Ecological Habitats on the Basis of Repetitive Extragenic Palindromic-PCR Fingerprinting. Appl. Environ. Microbiol. 70: 3149-3151 [Abstract] [Full Text]