Bielefeld sends a plasmid based on BBa_I746909 that has a translation enhancing sequence (5’-UTR), and Freiburg sends a plasmid containing turboYFP, a His- and a Halo-Tag. We would like to compare if these parts work in different cell-free proteins synthesis environments.

To test the plasmid BBa_I746909 containing a translation enhancing sequence (5'-UTR) we compared it to our GFPs used for cell-free expression (HA-GFP-His6-His6 and His-GFP-Spy). Both plasmids were treated alike and compared to a sample containing no DNA (negative control) and a dilution series of expressed and purified GFP (positive control). All reactions were performed in triplicates. The samples were expressed for 2 hours at 37°C in a 384-well plate using our own lysate and premix. After expression, a western blot and dot blot were performed.

The iGEM Team Stockholm wants to develop a bacterial biomarker for early detection of cancer biomarkers. We thought as we are both working on diagnostic tools it would be great to start a collaboration.

Therefore, we wanted to measure with our DiaChip the interaction of one of their affibodies to the Her2-protein, one of their cancer biomarkers. Team Stockholm expressed the affibody with a His-Tag, so it would bind to our Ni-NTA surface and sent the "E. coli" lysate to us. From R&D-Systems we reveived the purified Her2-antigen. Unfortunately, the antigen also had a His-Tag, so we couldn't measure it on Ni-NTA slides, because it would not only bind the affibody but also the Ni-NTA on the surface. This would lead to an increased optical thickness all over the slide and we would not be able to get a specific signal. If we had spotted the affibody lysate on an unspecific surface, all other proteins in the lysate would have bound to the surface as well. This would have resulted in an affibody concentration on the spot below the detection limit. So that wasn't a possibility either.

Our last chance was to spot the antigen on an unspecific surface and flush over the affibody lysate. Due to the small molecular weight of the affibody (10 kDa), it is really hard to see a binding in the iRIf (detection limit: ~10 kDa). Nevertheless we still tried.

We contributed to the iGEM Newsletter published by the iGEM Team Amoy.

The iGEM Team Amoy published regular Newsletters for the iGEM competition together with Paris_Bettencourt and Pasteur_Paris. It provided all iGEM Teams with the possibility to share their project idea, information about experiments they are performing or opinions about crucial topics of synthetic biology as well as to ask for help with complications they faced during summer. All in all, there were seven issues published with different contents, three of them being special issues dealing with setting up an iGEM Team, the current situation of synthetic biology and software used in iGEM.

We were asked if we would like to contribute to the work of the iGEM Team and used this opportunity to share our project and thoughts with the iGEM community. We were pleased to be able to contribute to such a great piece of work.

All the issues that were published during this year's iGEM competition and further information can be found here.

The iGEM Team Tübingen provided us with a Spy-tagged protein.

For the establishment of a specific surface on the glass slide of the DiaCHIP we followed different approaches. Among others, we planned to test a surface with a SpyCatcher immobilized on top of an activated silane layer. The SpyCatcher would then interact specifically with cell-free expressed Spy-tagged proteins. In order to be able to verify the predicted function of our surface before performing cell-free expression, the iGEM Team Tübingen kindly provided us with a purified carboxyfluorecine labelled SpyTag.

Unfortunately, due to time constraints, we were not able to establish a SpyCatcher surface and test the corresponding SpyTag we received from Team Tübingen.