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Open Access Methodology article

DNA-free RNA preparations from mycobacteria

Joachim Stephan1, Johannes G Bail1, Fritz Titgemeyer1 and Michael Niederweis12*

Author Affiliations

1 Lehrstuhl für Mikrobiologie, Friedrich-Alexander-Universität Erlangen-Nürnberg, Staudtstr. 5, D-91058 Erlangen, Germany

2 Department of Microbiology, University of Alabama at Birmingham, 609 Bevill Biomedical Research Building, 845 19th Street South, Birmingham, AL 35294, USA

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BMC Microbiology 2004, 4:45  doi:10.1186/1471-2180-4-45

Published: 30 November 2004

Abstract

Background

To understand mycobacterial pathogenesis analysis of gene expression by quantification of RNA levels becomes increasingly important. However, current preparation methods yield mycobacterial RNA that is contaminated with chromosomal DNA.

Results

After sonication of RNA samples from Mycobacterium smegmatis genomic DNA is efficiently removed by DNaseI in contrast to untreated samples.

Conclusions

This procedure eliminates one of the most prevalent error sources in quantification of RNA levels in mycobacteria.