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DNA Stable-Isotope Probing DNA-SIP
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Quantitative microbial ecology through stable isotope probing.

Bruce A Hungate1, Rebecca L Mau2, Egbert Schwartz3

  • 1Center for Ecosystem Science and Society, Northern Arizona University, Flagstaff, Arizona, USA Department of Biological Sciences, Northern Arizona University, Flagstaff, Arizona, USA Bruce.Hungate@nau.edu.

Applied and Environmental Microbiology
|August 23, 2015
PubMed
Summary
This summary is machine-generated.

Quantitative stable isotope probing (qSIP) measures bacterial isotope enrichment with high taxonomic resolution. This new method reveals significant variations in bacterial element cycling and substrate utilization in environmental samples.

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Area of Science:

  • Microbiology
  • Environmental Science
  • Biogeochemistry

Background:

  • Quantifying elemental transformation rates by bacteria in diverse environments is challenging.
  • Existing methods for determining isotope enrichment with fine taxonomic resolution are limited.
  • Stable isotope probing (SIP) offers potential but requires refinement for quantitative analysis.

Purpose of the Study:

  • To develop a modified stable isotope probing technique enabling quantitative measurement of isotope enrichment in individual bacterial taxa.
  • To introduce quantitative stable isotope probing (qSIP) for precise assessment of bacterial element assimilation.

Main Methods:

  • DNA is fractionated into multiple densities after isopycnic centrifugation.
  • Taxon-specific density curves are generated for labeled and unlabeled samples.
  • Density shifts are calculated relative to unlabeled controls to quantify isotope assimilation.

Main Results:

  • qSIP successfully quantified taxonomic variations in (18)O and (13)C enrichment in soil bacteria.
  • Glucose addition enhanced (18)O assimilation, indicating indirect stimulation of other substrate utilization.
  • Demonstrated the quantitative benefits of qSIP in revealing complex bacterial responses.

Conclusions:

  • qSIP provides a robust method for measuring quantitative isotope enrichment in bacterial taxa.
  • The technique enhances understanding of microbial element cycling and metabolic responses in situ.
  • qSIP advances the study of microbial ecology and biogeochemical processes.