Difference between revisions of "Team:USP-EEL-Brazil/InterLab"

Interlab

IGEM is an international Synthetic Biology competition, which aims to set up standard protocols that work in any laboratory on the planet. To manage this task, Interlab Study was created, inviting all teams to participate by performing the same experiment: making measures of the Green Fluorescent Protein (GFP) to determine the feasible cell population.

In order to determine cell population, scientists usually use spectrophotometry to measure cells culture absorbance in 600 nm, and through it, the optical density (OD) can be obtained. So, theoretically, the higher cells number the higher OD. However, the values of OD obtained are under high variability, because of different calibration and precision between labs equipment, different glassware size, and others lab-to-lab factors. In addition, through this methodology, it isn’t possible to separate dead feasible cells.

In this sense, to respond to all the problems above, the methodology of measuring GFP is another way to determine feasible cells, once the dead ones will not express GFP. Through the GFP measurement methodology, the Measurement Committee wants our help to answer the following question:

‘’Can we reduce lab-to-lab variability in fluorescence measurements by normalizing to absolute cell count or colony-forming units (CFUs) instead of OD?‘’

In order to achieve this goal, Team USP-EEL-Brazil followed the Interlab protocols and sent the results to iGEM's measurement committee.

Materials and Methods

In order to make the experiments, we followed the method in the link below: (colocar o link)

The plasmids used are available in the iGEM kit:

Equipment used for measurements: TECAN INFINITY M200 PRO Plate used for measurements: Corning^® Costar Clear Polystyrene 96-Well Plates

Results