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Neufeld, J.D.* ; Vohra, J.* ; Dumont, M.G.* ; Lüders, T. ; Manefield, M.* ; Friedrich, M.W.* ; Murrell, J.C.*

DNA stable-isotope probing.

Nat. Protoc. 2, 860-866 (2007)
DOI
Open Access Green möglich sobald Postprint bei der ZB eingereicht worden ist.
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 (13C) or nitrogen (15N) 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.
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Publikationstyp Artikel: Journalartikel
Dokumenttyp Wissenschaftlicher Artikel
ISSN (print) / ISBN 1754-2189
e-ISSN 1750-2799
Zeitschrift Nature Protocols
Quellenangaben Band: 2, Heft: 4, Seiten: 860-866 Artikelnummer: , Supplement: ,
Verlag Nature Publishing Group
Begutachtungsstatus Peer reviewed
Institut(e) Institute of Groundwater Ecology (IGOE)