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Proper experimental design requires randomization/balancing of molecular ecology experiments |
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| Authors: | Miklós Bálint Orsolya Márton Marlene Schatz Rolf‐Alexander Düring Hans‐Peter Grossart |
| Affiliation: | 1. Senckenberg Biodiversity and Climate Research Centre, Frankfurt am Main, Germany;2. Institute for Soil Sciences and Agricultural Chemistry, Centre for Agricultural Research, Hungarian Academy of Sciences, Budapest, Hungary;3. Institut für Bodenkunde und Bodenerhaltung, Gie?en, Germany;4. Leibniz Institute for Freshwater Ecology and Inland Fisheries, Stechlin, Germany;5. Institute of Biochemistry and Biology, Potsdam University, Potsdam, Germany |
| Abstract: | Properly designed (randomized and/or balanced) experiments are standard in ecological research. Molecular methods are increasingly used in ecology, but studies generally do not report the detailed design of sample processing in the laboratory. This may strongly influence the interpretability of results if the laboratory procedures do not account for the confounding effects of unexpected laboratory events. We demonstrate this with a simple experiment where unexpected differences in laboratory processing of samples would have biased results if randomization in DNA extraction and PCR steps do not provide safeguards. We emphasize the need for proper experimental design and reporting of the laboratory phase of molecular ecology research to ensure the reliability and interpretability of results. |
| Keywords: | batch effect bias DNA extraction environmental DNA laboratory practice lake community metabarcoding nondemonic intrusions PCR sediment |