Team:Aix-Marseille/Measurement

Chew fight

Interlab study results

Results

The Team AMU decided to study 5 different promoters from the collection in table 1 (J23117, J23106, J23118, J23101 and J23100) and compare also the T7 promoter Bba_K525998 that we used for some of our constructions (J23117, J23106, J23118, J23101 and J23100). We could then measure the strength of the promoter we used under standard conditions in a well characterized assay.Each promoter was integrated into a construction expressing GFP Bba_I13504 with a strong/weak RBS (BBa_B0034).

Measurement methods

We choose to use 4 different methods to study the strength of the different promoters:

- TECAN microplate reader
- Fluorometer (SPEX fluorlog)
- Magicbox (home made filter based fluorimeter)
- Confocal Microscope

Our machines

Fluorimeter

Magic box

TECAN

Magic box

Each machine (if possible) was calibrated using a standard solution of the dye Atto488. This allow us to compare the measurements on the different instruments. Unfortunately the confocal microscopy couldn’t be calibrated this way as with the solution there was nothing to see, no signal and impossible to focus the microscope on nothing!!

TECAN results

Fluorometer results

Magicbox results

Confocal microscopy results

Conclusion

The results show that the promoter J23100 is the strongest in all methods. In contrary the promoter J23117 is weaker than the other promoters. The promoter T7 hasn’t got basal expression detectable. The promoter’s order of the strength is little bit variable depends on the different methods.

	TECAN	Fluorometer	Magicbox	Confocal Microscopy
J23100	+++++	+++++	+++++	+++++
J23101	++++	++++	+++	+++++
J23118	++++	+++	+++++	+++++
J23106	++	++	+	+
J23117	+	+	+	+
P T7	-	-	-	-

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