Measurement Error and Resolution in Quantitative Stable Isotope Probing: Implications for Experimental Design
Ella T. Sieradzki, Benjamin J. Koch, Alex Greenlon, Rohan Sachdeva, Rex R. Malmstrom, Rebecca L. Mau, Steven J. Blazewicz, Mary K. Firestone, Kirsten Hofmockel, Egbert Schwartz, Bruce A. Hungate, Jennifer Pett‐Ridge
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.