Team:UCopenhagen/Safety
Calibrations
Human, animal and environmental safety considerations are prerequisite to commencing any legitimate science project. Working with genetically modified organisms involves advanced regulation owing to its political potency. The relatively recent emergence of synthetic biology and its revolutionary potential earns even more public scrutiny. Vigilant enforcement of safety best practices is critical to preventing avoidable public misconceptions.
Incell has received extensive safety training from the Department of Plant and Environmental Sciences at the University of Copenhagen. From the very beginning we integrated safety into the concept creation and experimental design. All decisions have been made in accordance with Danish, EU and WHO safety legislation. At Incell, safety is baked in.
Cell measurements
Two colonies from each transformation were picked, and grown in foil-covered 50 ml falcon tubes over night (18 hours).
Preparation OD was measured, and a dilution was calculated to achieve an OD of 0.02. Dilution calculations can be found in the table next to this. Here we used the calculated correction factor from our initial abs/OD calibration. From the absorption measurements taken at 0 hours, we see indications of pipetting errors, as the OD600 (average) ranges from 0.015 to 0.6 (table 1).
OD600
Cell growth stagnated between 4 and 6 hours. Cells transformed with Test Device 1 and 4 grew slower than the 6 other transformations, and even decreased in OD between 4 and 6 hours. Click on figures for enlarged images:
Fluorescence
Some transformed cells continue to increase fluorescence despite a decrease in OD in the same samples. Devices 3 and 6 are very close to the negative control in fluorescence. Click on figures for enlarged images:
Conclusion
Our data indicate which plasmid elements induce the highest production of GFP.
Plasmids containing J23117 (Test devices 3 and 6) does not express fluorescence to a higher degree than the negative control. Plasmids with J23101 (Device 1 and 4) induced the highest fluorescence, and plasmids containing J23106 (Test devices 2 and 5) were somewhere in between. Combining the J23101 or J23106 with I13504 (Test devices 1-3) gave a higher fluorescence than adding BCD2.E0040.B0015 (Test devices 4-6).
These results are not reliable on their own, but will be more robust and reliable when combined with data from the other teams participating in the interlab study.
