Design of Experiments (DoE) is a statistical method that allowed us to design the most efficient experiments to determine the factors that influence the expression of our PPK enzyme and the expression of our Eut (microcompartment) proteins. By using DoE, we can efficiently explore a very large experimental space in a small number of experiments, allowing us to test multiple hypotheses at once in a rapid and robust manner. Performing our measurements using the statistical tools of DoE allowed us to develop an improved understanding of the experimental factors affecting protein expression in the Phosphostore system, within the limited time frame available for an iGEM project.

Other variations of DoE allow thorough mapping of the experimental space, which would allow rapid characterisation of parts in a broad range of conditions. Thorough characterisation of parts using DoE-based methodology can allow accurate predictions of part behaviour in variable conditions.

We used DoE to optimise expression of a polyphosphate kinase construct: PduD(1-20)_mCherry_cgPPK
To see how we did this, look at or Design of Experiments section

We also used DoE to optimise expression of the Eut micro-compartment structural gene operons Tet_EutMN and araB_eutLK