Difference between revisions of "Team:Baltimore Bio-Crew/Results"

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{{Baltimore_Bio-Crew}}
 
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{{Team:Baltimore Bio-Crew/JS}}
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{{Team:Baltimore Bio-Crew/CSS}}
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{{Team:Baltimore Bio-Crew/templateReset}}
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<html>
 
<html>
  
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<style>
  
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<div class="column full_size" >
 
  
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<h1>Results</h1>
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article {
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font-size: 16px;
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font-style: italic;
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padding-top: 10px;
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padding-bottom: 10px;
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font-family: 'Abel', san-serif;
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color: black;
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width: 50%;
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margin:0 auto;
  
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<p>Here you can describe the results of your project and your future plans. </p>
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}
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.footerSection{
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background-color:#a5f7c3;
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      margin-top: 300px;
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}
  
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<h5>What should this page contain?</h5>
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footer{
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<ul>
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width:60%;
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<li> Clearly and objectively describe the results of your work.</li>
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margin-left:30%;
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<li> Future plans for the project. </li>
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}
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<li> Considerations for replicating the experiments. </li>
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</ul>
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<h5>You should also describe what your results mean: </h5>
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.html{
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  height: 100%;
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}
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.Intro{
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  width: 100%;
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  height:100%;
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  margin: 0 0 0 0;
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}
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h1{
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  font-size: 100px;
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  text-align: center;
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  color: black;
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  font-family: 'Saira', sans-serif;
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  padding-top: 40px;
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text-shadow: 4px 4px #3bb1e2;
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}
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h3{
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  font-size: 25px;
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  font-style: italic;
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  text-align: center;
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  padding-top: 20px;
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  padding-bottom: 30px;
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  font-family: 'Abel', san-serif;
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  font-style: italic;
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  color: black;
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text-decoration: underline;
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}
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h4{
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  font-size: 18px;
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  font-style: italic;
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  text-align: center;
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  padding-top: 20px;
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  padding-bottom: 30px;
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  font-family: 'Abel', san-serif;
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  color: black;
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}
  
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<ul>
 
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<li> Interpretation of the results obtained during your project. Don't just show a plot/figure/graph/other, tell us what you think the data means. This is an important part of your project that the judges will look for. </li>
 
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<li> Show data, but remember all measurement and characterization data must be on part pages in the Registry. </li>
 
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<li> Consider including an analysis summary section to discuss what your results mean. Judges like to read what you think your data means, beyond all the data you have acquired during your project. </li>
 
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</ul>
 
  
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h2{
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  padding: 75px 0 0 0;
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  font-size: 36px;
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  font-style: normal;
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  font-family: 'Saira', sans-serif;
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  text-align: center;
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color: black;
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}
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</style>
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      </head>
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      <body>
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<section>
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<header>
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        <div class="Intro">
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          <h1>BALTIMORE BIO-CREW</h1>
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          <h4>Bio-Engineering E.Coli To Degrade Plastic and Save The Baltimore Inner Harbor</h4>
 
</div>
 
</div>
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<hr>
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</header>
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</section>
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<section id="description" class= "projectDescription">
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<header>
  
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<div class="clear"></div>
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                        <h3>Results</h3>
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</header>
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<article>
  
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<div class="column half_size" >
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Working hard, we managed to successfully clone the Esterase gene into E.coli. Both colony PCR and sequencing were positive for the CE 5 esterase clone. However, Lipase was not successfully cloned and the sequences came up negative and truncated.</article>
 +
<article>
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<img src="https://static.igem.org/mediawiki/2017/e/ee/BaltimoreBiocrewFigure1.png" style="width:960px; height: 540px;"><!--original size: 1,920 × 1,080 pixels-->
 +
</article>
 +
<article>
 +
Due to the negative results with the lipase, we hypothesized that the lipase enzyme was toxic to the strain of E. coli we were using, causing it to reject the DNA.
 +
</article>
 +
<article>
 +
We started inducing the positive esterase clone to produce protein, and running the protein produced on a protein gel after purifying on a nickel column. After running multiple protein gels with the esterase, no positive results were found.
 +
</article>
 +
<article>
 +
Because of this, we hypothesized that the E. coli cells were not producing the esterase because the enzyme was also harmful to the cells.
 +
</article>
 +
<article>
 +
<img src="https://static.igem.org/mediawiki/2017/5/5e/T--Baltimore_Bio-Crew--results_fig2.jpg" style="width:960px; height: 540px;"><!--original size: 1,920 × 1,080 pixels-->
 +
</article>
 +
<article>
 +
To bypass the cell issue and test whether or not our constructs could actually produce the protein, we decided to test the constructs and biobricks on their own by translating them using a cell free kit, and then running the product of that on a protein gel. The protein gel results after the cell free kit were inconclusive, possibly because the cell just wasn’t producing enough protein to show up on the gel.
 +
</article>
 +
<article>
 +
<img src="https://static.igem.org/mediawiki/2017/e/ec/T--Baltimore_Bio-Crew--results_fig3.jpg" style="width:960px; height: 540px;"><!--original size: 1,920 × 1,080 pixels-->
 +
</article>
  
  
−
<h5> Project Achievements </h5>
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<article>
 +
We decided to run the cell free kit products through a FDA enzyme test. Fluorescence was seen, indicating that enzyme was being produced! The results of this test are shown on a bar graph below
 +
</article>
 +
<article>
 +
<img src="https://static.igem.org/mediawiki/2017/3/38/T--Baltimore_Bio-Crew--results_fig4.jpg" style="width:960px; height: 540px;"><!--original size: 1,920 × 1,080 pixels-->
 +
</article>
 +
<article>
 +
Additional tests will need to be done, but the FDA test indicated that our constructs can produce protein, even if the enzymes may not be able to be produced in E. coli
 +
</article>
  
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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>
 
  
−
</div>
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</section>
  
 +
<section id="Footer" class="footerSection">
 +
<hr>
 +
 +
<h2>
 +
Sponsors
 +
</h2>
 +
<h4>
 +
The Baltimore Bio-Crew thanks our sponsors for their generous support of our team that made our project and travel to the Jamboree possible. Thank you!
 +
</h4>
  
−
<div class="column half_size" >
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<footer>
  
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<h5>Inspiration</h5>
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<a href="http://www.bd.com/en-us">
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<p>See how other teams presented their results.</p>
+
  <img src="https://upload.wikimedia.org/wikipedia/en/f/f8/Update_Color_BD_PNG_Logo.png" alt="BD Medical Technology, Advancing the World of Health - BD" style="width:100px; height:100px;">
−
<ul>
+
</a>
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<li><a href="https://2014.igem.org/Team:TU_Darmstadt/Results/Pathway">2014 TU Darmstadt </a></li>
+
<a href="http://familyleague.org/">
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<li><a href="https://2014.igem.org/Team:Imperial/Results">2014 Imperial </a></li>
+
  <img src="http://baltimoreattendance.org/wp-content/uploads/2015/08/flbcinc-360x230.png" alt="Family League of Baltimore" style="width:100px; height:100px;">
−
<li><a href="https://2014.igem.org/Team:Paris_Bettencourt/Results">2014 Paris Bettencourt </a></li>
+
</a>
−
</ul>
+
  
−
</div>
+
                                      <a>
 +
  <img src="https://static.igem.org/mediawiki/2017/6/6c/T--Baltimore_Bio-Crew--fabian_kolker_small_icon.png" alt="Fabian Kolker Foundation" style="width:100px; height:100px;">
 +
</a>
  
 +
<a href="http://vwrfoundation.org/">
 +
  <img src="https://static.igem.org/mediawiki/2016/1/1a/T--Baltimore_Biocrew--VWR_Foundation_LOGO.jpeg" alt="VWR Charitable Foundation" style="width:100px; height:100px;">
 +
</a>
 +
<a href="http://www.marylandrecyclingnetwork.org/">
 +
  <img src="https://media.licdn.com/mpr/mpr/shrink_200_200/AAEAAQAAAAAAAAI8AAAAJDY0ZDg0ZjlkLWVlMTItNGI1Mi1iNWEwLWYzMDVlYWMwMTZhZg.png" alt="Maryland Recycling Network" style="width:100px; height:100px;">
 +
</a>
  
 +
<a href="https://www.rwdfoundation.org/">
 +
  <img src="https://static.igem.org/mediawiki/2016/6/65/T--Baltimore_BioCrew--DeutschFoundation.png" alt="The Robert W. Deutsch Foundation" style="width:100px; height:100px;">
 +
</a>
  
 +
</footer>
 +
</section>
 +
      </body>
 
</html>
 
</html>

Latest revision as of 19:40, 19 November 2017


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BALTIMORE BIO-CREW

Bio-Engineering E.Coli To Degrade Plastic and Save The Baltimore Inner Harbor

Results

Working hard, we managed to successfully clone the Esterase gene into E.coli. Both colony PCR and sequencing were positive for the CE 5 esterase clone. However, Lipase was not successfully cloned and the sequences came up negative and truncated.
Due to the negative results with the lipase, we hypothesized that the lipase enzyme was toxic to the strain of E. coli we were using, causing it to reject the DNA.
We started inducing the positive esterase clone to produce protein, and running the protein produced on a protein gel after purifying on a nickel column. After running multiple protein gels with the esterase, no positive results were found.
Because of this, we hypothesized that the E. coli cells were not producing the esterase because the enzyme was also harmful to the cells.
To bypass the cell issue and test whether or not our constructs could actually produce the protein, we decided to test the constructs and biobricks on their own by translating them using a cell free kit, and then running the product of that on a protein gel. The protein gel results after the cell free kit were inconclusive, possibly because the cell just wasn’t producing enough protein to show up on the gel.
We decided to run the cell free kit products through a FDA enzyme test. Fluorescence was seen, indicating that enzyme was being produced! The results of this test are shown on a bar graph below
Additional tests will need to be done, but the FDA test indicated that our constructs can produce protein, even if the enzymes may not be able to be produced in E. coli