This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Weinbauer, M. G.
Right arrow Articles by Höfle, M. G.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Weinbauer, M. G.
Right arrow Articles by Höfle, M. G.
Agricola
Right arrow Articles by Weinbauer, M. G.
Right arrow Articles by Höfle, M. G.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, March 2002, p. 1082-1087, Vol. 68, No. 3
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.3.1082-1087.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Simultaneous Extraction from Bacterioplankton of Total RNA and DNA Suitable for Quantitative Structure and Function Analyses

Markus G. Weinbauer,* Ingo Fritz, Dirk F. Wenderoth, and Manfred G. Höfle

Department of Environmental Microbiology, AG Microbial Ecology, GBF-German Research Center for Biotechnology, D-38100 Braunschweig, Germany

Received 22 May 2001/ Accepted 10 October 2001

The aim of this study was to develop a protocol for the simultaneous extraction from bacterioplankton of RNA and DNA suitable for quantitative molecular analysis. By using a combined mechanical and chemical extraction method, the highest RNA and DNA yield was obtained with sodium lauryl sarcosinate-phenol or DivoLab-phenol as the extraction mix. The efficiency of extraction of nucleic acids was comparatively high and varied only moderately in gram-negative bacterial isolates and bacterioplankton (RNA, 52 to 66%; DNA, 43 to 61%); significant amounts of nucleic acids were also obtained for a gram-positive bacterial isolate (RNA, 20 to 30%; DNA, 20 to 25%). Reverse transcription-PCR and PCR amplification products of fragments of 16S rRNA and its genes were obtained from all isolates and communities, indicating that the extracted nucleic acids were intact and pure enough for community structure analyses. By using single-strand conformation polymorphism of fragments of 16S rRNA and its gene, community fingerprints were obtained from pond bacterioplankton. mRNA transcripts encoding fragments of the enzyme nitrite reductase gene (nir gene) could be detected in a pond water sample, indicating that the extraction method is also suitable for studying gene expression. The extraction method presented yields nucleic acids that can be used to perform structural and functional studies of bacterioplankton communities from a single sample.

* Corresponding author. Present address: Department of Biological Oceanography, The Netherlands Institute for Sea Research, P.O. Box 59, 1970 AB Den Burg, Texel, The Netherlands. Phone: 31-222-369-539. Fax: 31-222-319-674. E-mail: wein{at}nioz.nl.

Applied and Environmental Microbiology, March 2002, p. 1082-1087, Vol. 68, No. 3
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.3.1082-1087.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Salka, I., Moulisova, V., Koblizek, M., Jost, G., Jurgens, K., Labrenz, M. (2008). Abundance, Depth Distribution, and Composition of Aerobic Bacteriochlorophyll a-Producing Bacteria in Four Basins of the Central Baltic Sea. Appl. Environ. Microbiol. 74: 4398-4404 [Abstract] [Full Text]  
  • Smith, C. J., Nedwell, D. B., Dong, L. F., Osborn, A. M. (2007). Diversity and Abundance of Nitrate Reductase Genes (narG and napA), Nitrite Reductase Genes (nirS and nrfA), and Their Transcripts in Estuarine Sediments. Appl. Environ. Microbiol. 73: 3612-3622 [Abstract] [Full Text]  
  • Kuiper, M. W., Valster, R. M., Wullings, B. A., Boonstra, H., Smidt, H., van der Kooij, D. (2006). Quantitative Detection of the Free-Living Amoeba Hartmannella vermiformis in Surface Water by Using Real-Time PCR. Appl. Environ. Microbiol. 72: 5750-5756 [Abstract] [Full Text]  
  • Eichler, S., Christen, R., Holtje, C., Westphal, P., Botel, J., Brettar, I., Mehling, A., Hofle, M. G. (2006). Composition and Dynamics of Bacterial Communities of a Drinking Water Supply System as Assessed by RNA- and DNA-Based 16S rRNA Gene Fingerprinting.. Appl. Environ. Microbiol. 72: 1858-1872 [Abstract] [Full Text]  
  • Brettar, I., Labrenz, M., Flavier, S., Botel, J., Kuosa, H., Christen, R., Hofle, M. G. (2006). Identification of a Thiomicrospira denitrificans-Like Epsilonproteobacterium as a Catalyst for Autotrophic Denitrification in the Central Baltic Sea. Appl. Environ. Microbiol. 72: 1364-1372 [Abstract] [Full Text]  
  • Labrenz, M., Jost, G., Pohl, C., Beckmann, S., Martens-Habbena, W., Jurgens, K. (2005). Impact of Different In Vitro Electron Donor/Acceptor Conditions on Potential Chemolithoautotrophic Communities from Marine Pelagic Redoxclines. Appl. Environ. Microbiol. 71: 6664-6672 [Abstract] [Full Text]  
  • Johnson, D. R., Lee, P. K. H., Holmes, V. F., Alvarez-Cohen, L. (2005). An Internal Reference Technique for Accurately Quantifying Specific mRNAs by Real-Time PCR with Application to the tceA Reductive Dehalogenase Gene. Appl. Environ. Microbiol. 71: 3866-3871 [Abstract] [Full Text]  
  • Labrenz, M., Brettar, I., Christen, R., Flavier, S., Botel, J., Hofle, M. G. (2004). Development and Application of a Real-Time PCR Approach for Quantification of Uncultured Bacteria in the Central Baltic Sea. Appl. Environ. Microbiol. 70: 4971-4979 [Abstract] [Full Text]  
  • Fey, A., Eichler, S., Flavier, S., Christen, R., Hofle, M. G., Guzman, C. A. (2004). Establishment of a Real-Time PCR-Based Approach for Accurate Quantification of Bacterial RNA Targets in Water, Using Salmonella as a Model Organism. Appl. Environ. Microbiol. 70: 3618-3623 [Abstract] [Full Text]  
  • Dominik, K., Hofle, M. G. (2002). Changes in Bacterioplankton Community Structure and Activity with Depth in a Eutrophic Lake as Revealed by 5S rRNA Analysis. Appl. Environ. Microbiol. 68: 3606-3613 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»