Difference between revisions of "Team:Wageningen UR/testpage"

(Undo revision 148047 by Lisaro (talk))
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{{Wageningen_UR/menu}}
 
{{Wageningen_UR/menu}}
 
<html>
 
<html>
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<!--Journal block-->
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<h1>Fungal resistance journal</h1>
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<p>All transformations were done in <i>E. coli</i>. Preparation of the backbones was done in DH5alfa, while for transformations with the clusters JM109 was used.</p>
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<div class="container" style="margin-top:100px;">
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<!--<h1>Lab journal</h1>-->
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<div id="timelineContainer" class="timelineContainer">
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<div class="timelineToggle">
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<p><a class="expandAll">+ expand all</a></p>
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</div>
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<br class="clear">
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<div class="timelineMajor">
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<h2 class="timelineMajorMarker"><span>May</span></h2>
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<dl class="timelineMinor">
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<dt id="02w1"><a>Growth experiments</a></dt>
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<dd class="timelineEvent" id="02w1EX" style="display:none;">
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<p>Growth experiments were conducted with fusaric acid concentrations ranging from 0 to 250 ug/ml were performed on <i>Eschericia coli (Strains DH5alfa, BL21 and JM109), Bacillus subtilis</i> and <i>Pseudomonas putida</i>. After 18 hours of incubation at 37 degress celcius, OD 600 measurements were taken.</p>
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                                                        <br class="clear">
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                                                        <figure>
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                                                        <img src="https://static.igem.org/mediawiki/2014/1/13/Wageningen_UR_journal_resistance_resistance_assay.png" width="800px">
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                                                        <figcaption>
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                                                        Figure 1. Both <i>B. subtilis</i> and all of the <i>E. coli</i> strains show a strong response to increasing doses of fusaric acid in the medium. <i>P. putida</i>, however, does not show any decrease in growth for the concentrations used. Therefore another experiment was done with higher concentrations for <i>P. putida</i>.
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                                                        </figcaption>
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                                                        </figure>
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                                                        <p><i><b>NOTE: After ordering new fusaric acid later in the project, it was found out that the fusaric acid used in this experiment was not the same strength, with the old one most likely being (partially) degraded. Therefore data from this experiment is not used.</b></i></p>
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                                                        </p>
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<br class="clear">
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</dd>
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<!-- /.timelineEvent -->
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</dl>
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<!-- /.timelineMinor -->
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</div>
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<!-- /.timelineMajor -->
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<div class="timelineMajor">
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<h2 class="timelineMajorMarker"><span>June</span></h2>
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<dl class="timelineMinor">
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<dt id="02w2"><a>HPLC</a></dt>
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<dd class="timelineEvent" id="02w2EX" style="display:none;">
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<p>Since some soil bacteria are known to be able to degrade fusaric acid (FA), a HPLC experiment was started to test <i>P. putida</i>(PP) for fusaric acid degradation and carbon utilization. M9 media was used with glucose/fusaric acid(250ug/ml) or both as a carbon source.  </p>
 +
                                                                                                             
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                                                        <p>For the HPLC the following settings were used.  </p>
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                                                        <p><ul>
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                                                        <li>Column: Polaris C18A</li>
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                                                        <li>Eluent: Acetonitril</li>
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                                                        <li>Temperature: 35 degrees Celsius</li>
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                                                        <li>Flow speed: 0.5 ml/min</li>
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                                                        <li>Detection: UV (260 - 280nm)</li>
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                                                        </ul></p>
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<br class="clear"/>
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                                                        <figure>
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                                                        <img src="https://static.igem.org/mediawiki/2014/c/cf/Wageningen_UR_journal_resistance_hplc_first_try.png" width="800px">
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                                                        <figcaption>
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                                                        Figure 2. Left: The HPLC method used delivered a good calibration curve, but peak resolvance was not optimal, with the FA peak having overlap with several smaller peaks in the samples with <i>P. putida</i>. Therefore it was decided to repeat the experiment to improve the HPLC method. </br>
 +
                                                        Right: Initial results on fusaric acid breakdown showed signs of breakdown, but because peak resolvance was not optimal and the measurements were not done in duplo more data was needed to draw conclusions.                                                     
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                                                        </figcaption>
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                                                        </figure>
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                                                        <p>Based on the initial results a more elaborate experiment was started in duplo. Six samples would be grown in duplo:</p>
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                                                        <ul>
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                                                        <li>A. Negative control for growth on M9 without carbon source. (PP+, Glucose-, FA-)</li>
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                                                        <li>B. Positive control for growth on M9 with carbon source (glucose).(PP+, Glucose+, FA-)</li>
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                                                        <li>C. Positive control for growth on M9 with carbon source and fusaric acid. (PP+, Glucose+, FA-)</li>
 +
                                                        <li>D. Test for PP growth on FA as carbon source.(PP+, Glucose-, FA+)</li>
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                                                        <li>E. E. Negative control for contamination and FA stability. (PP+, Glucose-, FA-)</li>
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                                                        <li>F. Negative control for contamination. (PP-, Glucose+, FA-)</li>
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                                                    </dd>
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<dt id="02w3"><a>Growth experiments</a></dt>
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<dd class="timelineEvent" id="02w3EX" style="display:none;">
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                                                        </dd>
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<br class="clear">
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</div>
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<!-- /#timelineContainer -->
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<br>
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<br>
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<section id="Bees">
 
<section id="Bees">
 
<h1><b>Saving the Bees</b></h1>
 
<h1><b>Saving the Bees</b></h1>
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<p>Lorem ipsum dolor sit amet, consectetur adipiscing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. Duis aute irure dolor in reprehenderit in voluptate velit esse cillum dolore eu fugiat nulla pariatur. Excepteur sint occaecat cupidatat non proident, sunt in culpa qui officia deserunt mollit anim id est laborum. </p></section>
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<h2>References</h2>
 
<h2>References</h2>
 
<ol class="references"> <!-- Use ol for numbered list, ul for bullet points-->
 
<ol class="references"> <!-- Use ol for numbered list, ul for bullet points-->

Revision as of 14:47, 3 October 2016

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Wageningen UR iGEM 2016

 

Fungal resistance journal

All transformations were done in E. coli. Preparation of the backbones was done in DH5alfa, while for transformations with the clusters JM109 was used.

+ expand all


May

Growth experiments

June

HPLC
Growth experiments



Saving the Bees

Lorem ipsum dolor sit amet, consectetur adipiscing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. Duis aute irure dolor in reprehenderit in voluptate velit esse cillum dolore eu fugiat nulla pariatur. Excepteur sint occaecat cupidatat non proident, sunt in culpa qui officia deserunt mollit anim id est laborum. iGEM team Wageningen 2016 has designed and created BeeT, a genetic system in Escherichia coli that detects the presence of Varroa. destructor in the bee hive and produces a combination of chitinases and Cry/Cyt/Mtx toxins to remove them! If you want to read more about the system and our work during the last six months, the iGEM team Wageningen 2016 invites you to continue surfing around our Wiki. 1

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Something

Text without p tags!

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Tiny Bee

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Tiny Bee

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References

    1. Evans, J. D., & Schwarz, R. S. (2011). Bees brought to their knees: microbes affecting honey bee health. Trends in microbiology, 19(12), 614-620 ↩