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Team:Copenhagen/Collaboration
DTU-2
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This year the danish DTU-2 iGEM Team has made a new standardized assembly method, called USER assembly. This assembly method does not need the use of restriction enzymes to cut the DNA, as the iGEM standard assembly method. Since some of our Cytochrome p450's had several illegal restriction sites according to the iGEM rules, we saw the opportunity to apply the DTU-2 assembly method to our project. This also gave the DTU team the possibility to test the USER assembly method on our parts. This initiated a strong collaboration between us two teams. Technical University of Denmark (where the DTU-2 team is situated) is geografically close to The University of Copenhagen, and we therefore have had ample opportunity to visit each other, ledding to sharing of laboratory experiences, challenges and success stories. During the course of our project, we experienced a huge problem, which was that we did not recieve the 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. It therefore seemed logical to try out the USER method on our CYP79 B1 and A2. The DTU-2 team received our physical DNA for the B1 and A2 and they modified these into USER assembly standardized parts. Once modified, these were used by the Copenhagen iGEM Team for an independent tryout of the USER assembly method, using the protocols set forth by the DTU-2 team. This turned out to be a huge success. 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 . |
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