Difference between revisions of "Team:Manchester/Model/DoE"

Achievements:

1. Expression optimisation of a new part

2. Robust characterisation of expression

3.

4.

Introduction

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.

Method - PduD(1-20)_mCherry_cgPPK Expression Optimization

In order to optimise the efficiency of our engineered phosphate-accumulating organism, we wanted to find the ideal expression conditions for the PduD(1-20)_mCherry_cgPPK. We used JMP software by SAS to design our experiments. We decided to focus our analysis on continuous factors as more information can be extracted from a small number of experiments. With this in mind, we chose to investigate the following factors:

• OD at induction
• IPTG concentration in inducer (induces
• Post induction

Results

Some nice graphs as those already in the presentation. More if possible. Prove that the only few iterations needed to get high optimisation

Conclusion

That this is a technique that works and should be used