Difference between revisions of "Team:Freiburg/Collaborations"
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<strong>Bielefeld</strong> sends a plasmid based on BBa_I746909 that has a translation enhancing sequence (5’-UTR), and <strong>Freiburg</strong> 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. | <strong>Bielefeld</strong> sends a plasmid based on BBa_I746909 that has a translation enhancing sequence (5’-UTR), and <strong>Freiburg</strong> 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. | ||
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<a class="media" href="https://static.igem.org/mediawiki/2015/f/f8/Freiburg_bielefeld_collab.jpg" title="bielefeld_collab.jpg"><img align="right" alt="" class="mediaright" src="https://static.igem.org/mediawiki/2015/f/f8/Freiburg_bielefeld_collab.jpg" width="200"/></a> | <a class="media" href="https://static.igem.org/mediawiki/2015/f/f8/Freiburg_bielefeld_collab.jpg" title="bielefeld_collab.jpg"><img align="right" alt="" class="mediaright" src="https://static.igem.org/mediawiki/2015/f/f8/Freiburg_bielefeld_collab.jpg" width="200"/></a> | ||
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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). | 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). | ||
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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. | 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. | ||
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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 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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Revision as of 19:42, 3 September 2015
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.
