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DNA stable-isotope probing
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Neufeld, Josh D., Vohra, Jyotsna, Dumont, Marc G., Lueders, Tillmann, Manefield, Mike, Friedrich, Michael W. and Murrell, J. C. (J. Colin) (2007) DNA stable-isotope probing. Nature, Volume 2 (Number 4). pp. 860-866. doi:10.1038/nprot.2007.109 ISSN 1754-2189.
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Official URL: http://dx.doi.org/10.1038/nprot.2007.109
Abstract
Stable-isotope probing is a method used in microbial ecology that provides a means by which specific functional groups of organisms that incorporate particular substrates are identified without the prerequisite of cultivation. Stable-isotope-labeled carbon (C-13) or nitrogen (N-15) sources are assimilated into microbial biomass of environmental samples. Separation and molecular analysis of labeled nucleic acids ( DNA or RNA) reveals phylogenetic and functional information about the microorganisms responsible for the metabolism of a particular substrate. Here, we highlight general guidelines for incubating environmental samples with labeled substrate and provide a detailed protocol for separating labeled DNA from unlabeled community DNA. The protocol includes a modification of existing published methods, which maximizes the recovery of labeled DNA from CsCl gradients. The separation of DNA and retrieval of unlabeled and labeled fractions can be performed in 4-5 days, with much of the time being committed to the ultracentrifugation step.
Item Type: | Journal Article | ||||
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Subjects: | Q Science > QD Chemistry | ||||
Divisions: | Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) > Biological Sciences ( -2010) | ||||
Journal or Publication Title: | Nature | ||||
Publisher: | Nature Publishing Group | ||||
ISSN: | 1754-2189 | ||||
Official Date: | 2007 | ||||
Dates: |
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Volume: | Volume 2 | ||||
Number: | Number 4 | ||||
Number of Pages: | 7 | ||||
Page Range: | pp. 860-866 | ||||
DOI: | 10.1038/nprot.2007.109 | ||||
Status: | Peer Reviewed | ||||
Publication Status: | Published |
Data sourced from Thomson Reuters' Web of Knowledge
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