DNA stable-isotope probing
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, 2 (4). pp. 860-866. ISSN 1754-2189Full text not available from this repository.
Official URL: http://dx.doi.org/10.1038/nprot.2007.109
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|
|Subjects:||Q Science > QD Chemistry|
|Journal or Publication Title:||Nature|
|Publisher:||NATURE PUBLISHING GROUP|
|Number of Pages:||7|
|Page Range:||pp. 860-866|
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