Difference between revisions of "Team:Pasteur Paris/Results"

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   <li>Quantification of the number of Plasmids in each bacteria.</li>
 
   <li>Quantification of the number of Plasmids in each bacteria.</li>
 
   <li>Determination of the best Promoter</li>
 
   <li>Determination of the best Promoter</li>
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<p>Here you can describe the results of your project and your future plans. </p>
 
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<h5>What should this page contain?</h5>
 
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<ul>
 
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<li> Clearly and objectively describe the results of your work.</li>
 
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<li> Future plans for the project </li>
 
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<li> Considerations for replicating the experiments </li>
 
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</ul>
 
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<h4> Project Achievements </h4>
 
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<p>You can also include a list of bullet points (and links) of the successes and failures you have had over your summer. It is a quick reference page for the judges to see what you achieved during your summer.</p>
 
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<ul>
 
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<li>A list of linked bullet points of the successful results during your project</li>
 
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<li>A list of linked bullet points of the unsuccessful results during your project. This is about being scientifically honest. If you worked on an area for a long time with no success, tell us so we know where you put your effort.</li>
 
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</ul>
 
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<h4>Inspiration</h4>
 
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<p>See how other teams presented their results.</p>
 
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<ul>
 
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<li><a href="https://2014.igem.org/Team:TU_Darmstadt/Results/Pathway">2014 TU Darmstadt </a></li>
 
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<li><a href="https://2014.igem.org/Team:Imperial/Results">2014 Imperial </a></li>
 
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<li><a href="https://2014.igem.org/Team:Paris_Bettencourt/Results">2014 Paris Bettencourt </a></li>
 
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</ul>
 
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<p><em>Gibson assembly:  </em></p>
 
<p><em>Gibson assembly:  </em></p>

Revision as of 09:55, 22 October 2015

Results

pNP-Assay

    Estimation of the activity of NB-Esterase, slowest enzyme of our degradation chain

    TPA toxicity

    • Assessment of the toxicity
    • Determination that TPA is not degraded

      Operon assembly

      • Succesful assembly of 2 of our 4 gene clusters (BBa_K1622000 and BBa_K1622001).
      • Optimization of DNA sequences for E.coli.

        • Interlab Study

      • Successful building of the 3 devices
      • Characterization of the 3 devices using a Tecan micro-plate reader.
      • Quantification of the number of Plasmids in each bacteria.
      • Determination of the best Promoter

        • Gibson assembly:

        - BioBricks submitted to be BioBrick registry:

        - Bba_K1622000: assembly of the Glycoaldehyde Dehydrogenase (BBa_K936011) and the Glycoaldehyde Reductase (BBa_K936023) in the plasmid PSB1C3 :

        - Bba_K1622001: assembly of the NB-Esterase (BBa_K808030) and the TPA transporteur (BBa_K808007)

         

        Interlab Study:

        - Transformation in DH5-alpha of each promoter (BBa_J23101, BBa_J23106, BBa_J23117) and the controls GFP (BBa_I13405) and WT.

        - qPCR of the transformed cells to determinate the plasmid copy number per strain.

        - fluorescence test of the GFP expression and determination of the fluorescence per plasmid.

         

        pNP assay:

        - Insertion of the NB-Esterase (BBa_K808030) in the plasmid pDG011

        - Transformation of BAP1 with the construct NB-Esterase (Bba_K808030) in the plasmid pDG011

        - Isolation of three clones who have our construct NB-Esterase (Bba_K808030) in the plasmid pDG011.

        - Demonstration of the degradation of the 4-NitroPhenyl Butyrate by our modified BAP1 with NB-Esterase in the pDG011 plasmid.

         

        TPA :

        - Test of the TPA toxicity thanks to a variation of the TPA concentration.

        - Solubilization of TPA and amelioration of the method.

         

         

        Problems:

        Gibson assembly: the exonuclease contained in the Master Mix uncovered the Lox Recombination sites because they were too close to the overlapping ends.

        Interlab Study:

        Different strain for qPCR and Fluorescence test/ Modeling:

        Because of the absence of experimental results, we can't model our enzymatic system.

        CRE: - the yeast assembly did not work. pNP assay: - Two of our tests are less decisive because we didn't wait for a long time to see a real difference enter control and modified bacteria. - transformation of the construct very hard.

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