Measurement Error and Resolution in Quantitative Stable Isotope Probing: Implications for Experimental Design

Author:

Sieradzki Ella T.1ORCID,Koch Benjamin J.23,Greenlon Alex1,Sachdeva Rohan4,Malmstrom Rex R.5,Mau Rebecca L.2,Blazewicz Steven J.6,Firestone Mary K.1,Hofmockel Kirsten S.78,Schwartz Egbert23,Hungate Bruce A.23ORCID,Pett-Ridge Jennifer6ORCID

Affiliation:

1. University of California Berkeley, Environmental Science and Policy Management, Berkeley, California, USA

2. Center for Ecosystem Science and Society, Northern Arizona University, Flagstaff, Arizona, USA

3. Department of Biological Sciences, Northern Arizona University, Flagstaff, Arizona, USA

4. University of California Berkeley, Earth and Planetary Sciences, Berkeley, California, USA

5. Department of Energy Joint Genome Institute, Berkeley, California, USA

6. Physical and Life Sciences Directorate, Lawrence Livermore National Laboratory, Livermore, California, USA

7. Pacific Northwest National Laboratory, Richland, Washington, USA

8. Department of Ecology, Evolution and Organismal Biology, Iowa State University, Ames, Iowa, USA

Abstract

One of the biggest challenges in microbial ecology is correlating the identity of microorganisms with the roles they fulfill in natural environmental systems. Studies of microbes in pure culture reveal much about their genomic content and potential functions but may not reflect an organism’s activity within its natural community. Culture-independent studies supply a community-wide view of composition and function in the context of community interactions but often fail to link the two. Quantitative stable isotope probing (qSIP) is a method that can link the identity and functional activity of specific microbes within a naturally occurring community. Here, we explore how the resolution of density gradient fractionation affects the error and precision of qSIP results, how they may be improved via additional experimental replication, and discuss cost-benefit balanced scenarios for SIP experimental design.

Funder

U.S. Department of Energy

Publisher

American Society for Microbiology

Subject

Computer Science Applications,Genetics,Molecular Biology,Modeling and Simulation,Ecology, Evolution, Behavior and Systematics,Biochemistry,Physiology,Microbiology

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