Team:Freiburg/Collaborations

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Collaboration with iGEM team Bielefeld

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.

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