Analyzing microorganisms in environmental samples using stable isotope probing with H218O

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Abstract

When analyzing environmental samples of microorganisms by stable isotope probing (SIP), labeling the DNA with H218O, instead of organic or nitrogenous compounds, offers important advantages because water cannot be used as an energy, carbon, or nitrogen source. As a result, addition of the label is unlikely to influence microbial growth rates in soil directly, and microbial communities can be exposed to the label for long periods of time because they are not exposed to abnormally high substrate concentrations. Because all organisms incorporate water into their DNA, performing SIP with H2 18O is a method for identifying microorganisms that have grown during incubation with H218O, as well as microorganisms that have not grown (i.e., did not incorporate the label) but survived the incubation. This protocol describes the use of SIP with H218O to study soil microorganisms.

Original languageEnglish (US)
JournalCold Spring Harbor Protocols
Volume4
Issue number12
DOIs
StatePublished - 2009

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Isotopes
Microorganisms
Soil
Isotope Labeling
Labels
Water
DNA
Nitrogen
Carbon
Soils
Growth
Labeling
Substrates

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

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abstract = "When analyzing environmental samples of microorganisms by stable isotope probing (SIP), labeling the DNA with H218O, instead of organic or nitrogenous compounds, offers important advantages because water cannot be used as an energy, carbon, or nitrogen source. As a result, addition of the label is unlikely to influence microbial growth rates in soil directly, and microbial communities can be exposed to the label for long periods of time because they are not exposed to abnormally high substrate concentrations. Because all organisms incorporate water into their DNA, performing SIP with H2 18O is a method for identifying microorganisms that have grown during incubation with H218O, as well as microorganisms that have not grown (i.e., did not incorporate the label) but survived the incubation. This protocol describes the use of SIP with H218O to study soil microorganisms.",
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AB - When analyzing environmental samples of microorganisms by stable isotope probing (SIP), labeling the DNA with H218O, instead of organic or nitrogenous compounds, offers important advantages because water cannot be used as an energy, carbon, or nitrogen source. As a result, addition of the label is unlikely to influence microbial growth rates in soil directly, and microbial communities can be exposed to the label for long periods of time because they are not exposed to abnormally high substrate concentrations. Because all organisms incorporate water into their DNA, performing SIP with H2 18O is a method for identifying microorganisms that have grown during incubation with H218O, as well as microorganisms that have not grown (i.e., did not incorporate the label) but survived the incubation. This protocol describes the use of SIP with H218O to study soil microorganisms.

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