Stable Isotope Probing in Environmental Microbiology Studies
S. Jane Fowler and Lisa M. Gieg
from: Applications of Molecular Microbiological Methods (Edited by: Torben L. Skovhus, Sean M. Caffrey and Casey R.J. Hubert). Caister Academic Press, U.K. (2014)
Traditional microbiological methods involving the isolation of microbes from the environment in pure culture have been shown to be ineffective at accessing the majority of microbial diversity. Methods that allow the study of microbes in their native environment or in mixed cultures have been gaining in popularity in recent years. Stable isotope probing is a method that allows the identification of the taxonomic groups that are actively metabolizing a substrate, typically in enriched cultures but also in in situ communities. This technique involves incubations with an isotopically- labelled substrate (e.g. 13C-labelled) during which time the isotope label is incorporated into the biomolecules of organisms actively degrading the substrate. This is followed by the extraction and analysis of these biomolecules in order to identify the organisms incorporating the isotope label. Stable isotope probing has been refined for the analysis of all classes of biomolecules, and has been used to identify the key microbes involved in a number of biological processes read more ...