Difference between revisions of "Team:DTU-Denmark/Achievements"

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       <p>The following overview highlights our&nbsp;achievements:</p>
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       <p>We would be lying, if we stated everything worked the first time. We made hundreds&nbsp;of agar plates, replica plated thousands of plates, and we ran the same PCR many times before finding the right annealing temperature. But that being said, we have learned from our mistakes and &nbsp;it has helped us connect as a team. We are therefore proud to say:</p>
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<li><strong>MAGE Subtilis</strong>
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<li>We established a platform for MAGE in&nbsp;<em>Bacillus subtilis.</em></li>
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<li>We engineered a NRPS using MAGE in <em>Bacillus subtilis</em>&nbsp;to make a new product.</li>
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<li>We tested the first generation of our high throughput screening.</li>
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<li><strong>Knock out of upp and amyE</strong>
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<li>We improved characterization of a BioBrick and explored uses of inteins for production of short synthetic peptides.</li>
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<li><strong>​</strong>Identified&nbsp;that <em>upp</em> and <em>amyE</em> are not optimal selection targets for knockout mutations.</li>
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<li>We were not able to duplicate&nbsp;<em>Sun2015 and</em>&nbsp;knock&nbsp;out the upp locus using 2kb ssDNA using&nbsp;oligos.</li>
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<li><b>Growth experiment for mutants</b>
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<li><b>​</b>We&nbsp;identified that the growth rate of the B. <em>subtilis </em>with a recombinase grew faster than the wild type. This is conflicting with the results by <em>Sun2015</em>&nbsp;</li>
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<li>Identified that the mismatch-repeare knockout did not change&nbsp;growth rate.</li>
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<li><strong>Protocol for multiple MAGE cycles in B.&nbsp;<em>subtilis </em>168</strong>
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<li><strong>​</strong>Quantified&nbsp;colony picking to be&nbsp;an ineffective method of quantifying MAGE frequency.</li>
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<li>Showed that the knockout of mutS has a profound&nbsp;efffect on the transformation frequency compared to the wildtype.</li>
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<li><strong>Optimal amount of oligo for MAGE</strong>
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<li>The&nbsp;optimal amount of oligo was identified.&nbsp;</li>
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<li><strong>OD calculator = Dilution calculator</strong>
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<li><strong>​</strong>We made a&nbsp;calculator that could compute the appropriate dilution from the OD600 measured.</li>
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Revision as of 09:16, 2 October 2015


Key Achievements

Scroll down for more

Key Achievements

We would be lying, if we stated everything worked the first time. We made hundreds of agar plates, replica plated thousands of plates, and we ran the same PCR many times before finding the right annealing temperature. But that being said, we have learned from our mistakes and  it has helped us connect as a team. We are therefore proud to say:

  • We established a platform for MAGE in Bacillus subtilis.
  • We engineered a NRPS using MAGE in Bacillus subtilis to make a new product.
  • We tested the first generation of our high throughput screening.
  • We improved characterization of a BioBrick and explored uses of inteins for production of short synthetic peptides.

 

Technical University of Denmark
Department of Systems Biology
Søltofts Plads 221
2800 Kgs. Lyngby
Denmark
P: +45 45 25 25 25
M: dtu-igem-2015@googlegroups.com