Team:UrbanTundra Edmonton/InterLab
InterLab
Background
This year, our team participated in the fourth InterLab Measurement Study which aimed to establish and test a standardized protocol to measure GFP that can produce common and consistent numbers from all across the world. Doing our part, we used a plate reader, which we were given access to from the UAlberta iGEM team along with support from members of the UAlberta iGEM team to measure GFP fluorescence. Our team followed the protocols outlined in the iGEM InterLab page.
Materials
Plasmids used:
Devices (from InterLab Measurement Kit):
Positive control
Negative control
Test Device 1: J23101+I13504
Test Device 2: J23106+I13504
Test Device 3: J23117+I13504
Test Device 4: J23101.BCD2.E0040.B0015
Test Device 5: J23106.BCD2.E0040.B0015
Test Device 6: J23117.BCD2.E0040.B0015
Strain used:
Competent cells (Escherichia coli strain DH5α)
Materials used:
LB (Luria Bertani) media
Chloramphenicol (stock concentration 25 mg/mL dissolved in EtOH - working stock 25 ug/mL)
50 ml Falcon tube (covered in foil to block light)
Incubator at 37°C
1.5 ml eppendorf tubes for sample storage
Ice bucket with ice
Pipettes
96 well plate, black with flat, transparent/clear bottom preferred (provided by UAlberta team)
Plate reader used:
Tecan SAFIRE II Multi-mode Plate Reader
Protocol
Discussion
Looking at the data curves of our graphs, there is a linear relation at low concentrations of fluorescein but as the concentration increases, the curve plateaus. This suggests that our detector may have been oversaturated.
