Team:UCopenhagen/Collaborations

C O L L A B O R A T I O N S

Collaboration is a powerful driver of progress in science. To stand on the shoulders of giants is increasingly unrealistic in a globalised world where scientists work in ever more specialised areas and without geographic or language barriers.

For science to continue to benefit from globalisation, we must advance the axiom and seek to build a human pyramid from the shoulders before and around us. Collaboration and open source data are at the heart of iGEM and so too the heart of Incell.

Below we describe our most significant collaborations within the iGEM community. The symbiosis we strive for in our project has shone brightly through the partnerships established with teams around the world. Our story — and the stories of our friends — have been mutually shaped by the diversity of our shared experiences, knowledge and perspectives.

Figure 1 Standard curve of fluorescein fluorescence. Fluorescence in arbitraty units (a.u.), fluorescein concentration in µM.

Introduction

We participated in InterLitab, as we want to be contribute to the scientific progress made through this globe spanning project. In InterLab, 6 test devices are inserted in E.coli D5 α, and the growth and fluorescence is measured.

We used the following plasmids provided by iGEM HQ to transform E.coli:

  • 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

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:

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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:

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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.

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