Team:Aix-Marseille/Measurement
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
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 | - | - | - | - |