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From 2011.igem.org

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Revision as of 16:11, 17 September 2011

Collaboration with DTU-2
This year the danish DTU-2 iGEM Team has made a new standardized assembly method, called USER assembly. Since some of our Cytochrome p450's had several illegal restriction sites according to the iGEM rules, we saw the opportunity to apply their method to our assembly process, which also gave them the possibility to test the method on our parts. This is the background for the collaboration that we initiated.

Since DTU is situated close to The University of Copenhagen we have had ample opportunity to visit each other. This has led to sharing of laboratory experiences, challenges and success stories.

Our huge problem was that we did not receive our sequence information on the human Cytochrome p450 DNA that was kindly sent to us from Dr. Guengerich from Vanderbilt University in Nashville. This sadly put our project to a halt. Half-way through our project we decided to get the DNA sequenced on our own. At this point we were very pressed for time though. As the USER assembly method is very fast compared to the standard iGEM assembly method, collaborating with DTU-2 allowed us to work with and move on with this part of the project. Alas their help came too late as we did not have the time to obtain the results we would have liked.

We also decided that it would be helpful to have the DTU-2 team try out their method on our Cyp79's. So we gave them some of coding DNA and the sequences for the cyp79 B1 and A2 and had them assemble the parts for us. This turned out to be a good idea as the Cyp79B1 was expressed only when we used the USER assembly method in contrast to reapeated failures when using the iGEM standard assembly method.

During collaboration we have realized that the USER assembly method has many advantages compared to the standard assembly method of molecular biology.

It is:
-Mutationless
-Simple
-Fast
-Reliable