Over the past three years, iGEM has advanced the frontiers of science with the two biggest interlaboratory studies ever done in synthetic biology. These studies established a baseline for replicability of fluorescence measurements and identified likely key sources of error, and have now been published as an open-access journal article in PLOS ONE.
The 2017 iGEM Interlab Study
The aim of this year’s Interlab Study is to establish a GFP measurement protocol based on engineering principles that anyone with a plate reader can use in their lab. Teams were once more asked to use the same exact protocol around the world to produce common, comparable units for measuring GFP with different plate readers. Teams who had access to flow cytometry systems were also encouraged to make flow cytometry measurements, namely counting cells.
Materials and methods
The 8 plasmids (Positive Control, Negative Control, Test Device 1, Test Device 2, Test Device 3, Test Device 4, Test Device 5, and Test Device 6 - locations listed below) from Kit Plate 6 were transformed into E. coli DH5-alpha cells. We used this transformation protocol
The machines we used are given below. Using these machines was a great opportunity to discover laboratories and people at the Institut Pasteur, and to learn new biological methods.
-Incubator: Unitron Infors AGCH-4103 Bottmingen
-Plate reader: VaroSkan Lux Thermo Scientific
-Flow cytometer : Cytoflex S Beckman Coulter
Results
We conducted both the experiments with the plate reader and the flow cytometer. The results can be downloaded with the links below:
Acknowledgements
The members of our team who conducted the Interlab Study experiments were not at all familiar with the equipment used. Thus, nothing would have been possible without the precious help of a few researchers at the Pasteur Institut. We would like to thank warmly:
- Dr Sandrine Schmutz for giving us access to the flow cytometer and teaching us how to use it.
- Pierre-Henri Commere platform engineer who also taught us how to use and work on the flow cytometer
- Finally Maxime Chazal for explaining to us how to use the plate reader
