Proper experimental design requires randomization/balancing of molecular ecology experiments

Ecol Evol. 2018 Jan 10;8(3):1786-1793. doi: 10.1002/ece3.3687. eCollection 2018 Feb.

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: DNA extraction; PCR; batch effect; bias; environmental DNA; laboratory practice; lake community; metabarcoding; nondemonic intrusions; sediment.

Associated data

  • figshare/10.6084/m9.figshare.4579681