Reproducibility of High-Throughput Plate-Reader Experiments in Synthetic Biology

Plate-reader assays are commonly conducted to quantify the performance of synthetic biological systems. However, on the basis of a survey of 100 publications, we find that most publications do not report critical experimental settings of plate reader assays, suggesting widespread issues in their reproducibility. Specifically, critical plate reader settings, including shaking time and covering method, either vary between laboratories or are not reported by the publications. Here, we demonstrate that the settings of plate reader assays have a significant impact on bacterial growth, recombinant gene expression, and biofilm formation. Furthermore, we show that the plate reader settings affect the apparent activity, sensitivity, and chemical kinetics of synthetic constructs, as well as alter the apparent effectiveness of antibiotics. Our results suggest the critical need for consistent reporting of plate reader protocols to ensure the reproducibility of the protocols. In addition, our work provides data for the setup of plate reader protocols in synthetic biology experiments.