Difference between revisions of "Team:elan vital korea/Result"

 
(7 intermediate revisions by 2 users not shown)
Line 9: Line 9:
 
     // Cache the Window object
 
     // Cache the Window object
 
     $window = $(window);
 
     $window = $(window);
               
+
 
 
   $('section[data-type="background"]').each(function(){
 
   $('section[data-type="background"]').each(function(){
 
     var $bgobj = $(this); // assigning the object
 
     var $bgobj = $(this); // assigning the object
                   
+
 
 
       $(window).scroll(function() {
 
       $(window).scroll(function() {
                   
+
 
 
         // Scroll the background at var speed
 
         // Scroll the background at var speed
         // the yPos is a negative value because we're scrolling it UP!                              
+
         // the yPos is a negative value because we're scrolling it UP!
 
         var yPos = -($window.scrollTop() / $bgobj.data('speed'));
 
         var yPos = -($window.scrollTop() / $bgobj.data('speed'));
       
+
 
 
         // Put together our final background position
 
         // Put together our final background position
 
         var coords = '50% '+ yPos + 'px';
 
         var coords = '50% '+ yPos + 'px';
Line 24: Line 24:
 
         // Move the background
 
         // Move the background
 
         $bgobj.css({ backgroundPosition: coords });
 
         $bgobj.css({ backgroundPosition: coords });
       
+
 
 
}); // window scroll Ends
 
}); // window scroll Ends
  
  });  
+
  });
  
 
});
 
});
Line 51: Line 51:
 
     background: url(https://static.igem.org/mediawiki/2015/8/87/ElanVitalKorea_bg_3.png) 50% 0 no-repeat fixed;
 
     background: url(https://static.igem.org/mediawiki/2015/8/87/ElanVitalKorea_bg_3.png) 50% 0 no-repeat fixed;
 
min-height:1000px;
 
min-height:1000px;
     height: 1000px;
+
     height: 1000px;
 
     margin: 0 auto;
 
     margin: 0 auto;
 
     width: 100%;
 
     width: 100%;
Line 61: Line 61:
 
     background: url(https://static.igem.org/mediawiki/2015/9/9c/Lightbg_copy.jpg) 50% 0 no-repeat fixed;
 
     background: url(https://static.igem.org/mediawiki/2015/9/9c/Lightbg_copy.jpg) 50% 0 no-repeat fixed;
 
     min-height:1000px;
 
     min-height:1000px;
height: auto;
+
height: 1000px;
margin: 0 auto;
+
    margin: 0 auto;
 
     width: 100%;
 
     width: 100%;
 
     max-width: 1920px;
 
     max-width: 1920px;
Line 70: Line 70:
 
#maintext {
 
#maintext {
 
     background: url(https://static.igem.org/mediawiki/2015/9/9c/Lightbg_copy.jpg) 50% 0 no-repeat fixed;
 
     background: url(https://static.igem.org/mediawiki/2015/9/9c/Lightbg_copy.jpg) 50% 0 no-repeat fixed;
     height: auto;
+
     height: 1000px;
 
min-height:1000px;
 
min-height:1000px;
 
     margin: 0 auto;
 
     margin: 0 auto;
Line 81: Line 81:
 
#maintext2 {
 
#maintext2 {
 
     background: url(https://static.igem.org/mediawiki/2015/9/9c/Lightbg_copy.jpg) 50% 0 no-repeat fixed;
 
     background: url(https://static.igem.org/mediawiki/2015/9/9c/Lightbg_copy.jpg) 50% 0 no-repeat fixed;
height: auto;
+
    height: 1000px;
 
min-height:1000px;
 
min-height:1000px;
 
     margin: 0 auto;
 
     margin: 0 auto;
Line 109: Line 109:
 
     color: #797478;
 
     color: #797478;
 
     font: 10px/1.5 Verdana, Helvetica, sans-serif;
 
     font: 10px/1.5 Verdana, Helvetica, sans-serif;
     float: left;    
+
     float: left;
 
     width: 16%;
 
     width: 16%;
 
     margin:  2% 2% 50px 2%;
 
     margin:  2% 2% 50px 2%;
 
}
 
}
+
 
 
.galleryItem h2 {
 
.galleryItem h2 {
 
     text-transform: uppercase;
 
     text-transform: uppercase;
 
}
 
}
+
 
 
.galleryItem img {
 
.galleryItem img {
 
     max-width: 100%;
 
     max-width: 100%;
Line 209: Line 209:
 
margin-left:20%;
 
margin-left:20%;
 
}
 
}
   
+
 
 +
figure {
 +
  display: block;
 +
  margin-before: 1em;
 +
  margin-after: 1em;
 +
  margin-start: 40px;
 +
  margin-end: 40px;
 +
}
 +
 
 +
figcaption {
 +
  padding: 5px;
 +
/*  font-family: 'Cherry Swash', cursive; */
 +
  font-size: 20px;
 +
  font-weight: 700;
 +
  border: none;
 +
  background: transparent;
 +
  word-wrap:normal;
 +
  text-align: center;
 +
}
 +
 
 +
figure img {
 +
  border: 1px solid #323232;
 +
  padding: 5px;
 +
  background: #fff;
 +
  border-radius: 12px;
 +
  box-shadow: 3px 3px 5px rgba(50, 50, 50, .5);
 +
}
  
 
</style>
 
</style>
Line 247: Line 273:
 
             </h5>
 
             </h5>
 
                         <br><br>
 
                         <br><br>
 
<p style="text-align:center;">
 
<video controls>
 
<source src="https://static.igem.org/mediawiki/2015/8/89/PlasmidConstructionProcess.mp4" type="video/mp4">
 
</video></p>
 
<br /><br />
 
  
 
<div class="inner">
 
<div class="inner">
 
                     <P style="text-align:left;">
 
                     <P style="text-align:left;">
 
                         <font color="black">
 
                         <font color="black">
Our project had two main stages: in the first stage, we made the relevant plasmids, and in the second stage, we tested the cells transformed with the resulting plasmids. On the second stage, we used control experiments to make
 
sure that the results we got was our actual results. For the control tests, we used
 
untransformed E. coli and pure AHL as control groups. <br><br>
 
  
Through the experiments, we confirmed the successful recombination of the test  
+
Our project had two main stages: in the first stage, we made the relevant plasmids, and in the second stage, we tested the cells transformed with the resulting plasmids. On the second stage, we used control experiments to make sure that the results we got was our actual results. For the control tests, we used untransformed E. coli and pure AHL as control groups.<br><br>
plasmid and the reporter plasmid, and the successful transformation of the test  
+
 
cell and the reporter cell. To make sure everything goes as planned, we tested  
+
Through the experiments, we confirmed the successful recombination of the test plasmid and the reporter plasmid, and the successful transformation of the test cell and the reporter cell. To make sure everything goes as planned, we tested the plasmids not only at the end, but also after the addition of each part.<br><br>
the plasmids not only at the end, but also after the addition of each part.<br><br>
+
 
 +
For our project, we engineered two main plasmids: the reporter plasmid and the test plasmid. The reporter cell, which are competent E. coli transformed by reporter plasmids, produces LuxR, and has an inducible promoter that is activated by a LuxR-AHL complex. This promoter activates the transcription of GFP, which is easily detectable. The test cell, which are competent E. coli transformed by test plasmids, produces lactonase, which breaks down AHL. For the test plasmid, we tried two different types of enzymes: AiiA, and LacZ or beta-galactosidase.<br><br>
 +
When the test cell is treated with AHL, it breaks down AHL. When the reporter cell is added to the mix, no GFP is produced, as there is no AHL, whereas GFP is produced when a control group is used instead of a test cell.<br><br>
  
For our project, we engineered two main plasmids: the reporter plasmid and the
 
test plasmid. The reporter cell, which are competent E. coli transformed by
 
reporter plasmids, produces LuxR, and has an inducible promoter that is
 
activated by a LuxR-AHL complex. This promoter activates the transcription of
 
GFP, which is easily detectable. The test cell, which are competent E. coli
 
transformed by test plasmids, produces lactonase, which breaks down AHL. For
 
the test plasmid, we tried two different types of enzymes: AiiA, and LacZ or
 
beta-galactosidase.. <br><br>
 
  
When the test cell is treated with AHL, it breaks down AHL. When the reporter
 
cell is added to the mix, no GFP is produced, as there is no AHL, whereas GFP
 
is produced when a control group is used instead of a test cell. <br><br>
 
  
 
We used the plasmids J61100, C0060, C0062, K823017, J37032, R0062, I732006
 
We used the plasmids J61100, C0060, C0062, K823017, J37032, R0062, I732006
We transformed and raised the cells in solid plates to observe growth. <br><br>
+
We transformed and raised the cells in solid plates to observe growth.<br><br>
 +
</font></p>
 +
 
 +
<figure>
 +
<img class="displayed" src="https://static.igem.org/mediawiki/2015/4/48/C0062.png">
 +
<figcaption>Picture A: C0062</figcaption>
 +
</figure>
 +
<br /><br />
 +
 
 +
<figure>
 +
<img class="displayed" src="https://static.igem.org/mediawiki/2015/4/41/C0060.png">
 +
<figcaption>Picture B: C0060</figcaption>
 +
</figure>
 +
<br /><br />
 +
 
 +
<figure>
 +
<img class="displayed" src="https://static.igem.org/mediawiki/2015/c/c5/I732006.png">
 +
<figcaption>Picture C: I732006</figcaption>
 +
</figure>
 +
<br /><br />
 +
 
 +
<figure>
 +
<img class="displayed" src="https://static.igem.org/mediawiki/2015/2/22/R0062.png">
 +
<figcaption>Picture D: R0062</figcaption>
 +
</figure>
 +
<br /><br />
 +
 
 +
<figure>
 +
<img class="displayed" src="https://static.igem.org/mediawiki/2015/1/1b/J37032.png">
 +
<figcaption>Picture E: J37032</figcaption>
 +
</figure>
 +
<br /><br />
 +
 
 +
<figure>
 +
<img class="displayed" src="https://static.igem.org/mediawiki/2015/1/1a/K823017.png">
 +
<figcaption>Picture F: K823017</figcaption>
 +
<br /><br />
 +
 
 +
<P style="text-align:left;">
 +
    <font color="black">
 +
We went through miniprep, then electrophoresis to confirm that the plasmids were the ones we wanted.<br><br>
 +
 
 +
For our initial electrophoresis results for the individual plasmids, look at:
 +
Picture H, Picture I, Picture J, Picture K Lane 8, Picture L Lane 5, Picture M Lane 1.<br><br>
 +
 
 +
After we confirmed that all the plasmids were as they should be, we started the recombination process.<br><br>
 +
We also checked on every step of the recombination process by electrophoresis.<br><br>
 +
</font></p>
 +
 
 +
<figure>
 +
<img class="displayed" src="https://static.igem.org/mediawiki/2015/4/45/Ladder_reference.png">
 +
<figcaption>Picture G: Ladder reference</figcaption>
 +
</figure>
 +
<br /><br />
 +
 
 +
<figure>
 +
<img class="displayed" src="https://static.igem.org/mediawiki/2015/d/d8/Lanes_1-8.png">
 +
<figcaption>Picture H: Lane 1 : DNA ladder marker,<br />
 +
Lane 3 : C0060+EcoRI+PstI,<br />
 +
Lane 6 : I732073+EcoRI+PstI,<br />
 +
Lane 7 : J37032+EcoRI+PstI,<br />
 +
Lane 8 : K823017+EcoRI+PstI</figcaption>
 +
</figure>
 +
<br /><br />
 +
 
 +
<figure>
 +
<img class="displayed" src="https://static.igem.org/mediawiki/2015/1/1a/Lanes_1-9.png">
 +
<figcaption>Picture I: Lane 1-3: C0062+EcoRI+PstI,<br />
 +
Lane 9: DNA Ladder Marker</figcaption>
 +
</figure>
 +
<br /><br />
 +
 
 +
<figure>
 +
<img class="displayed" src="https://static.igem.org/mediawiki/2015/4/4b/Lanes_1-3.png">
 +
<figcaption>Picture J: Lane 1: DNA Ladder Marker,<br />
 +
Lane 2-3: R0062+EcoRI+SpeI</figcaption>
 +
</figure>
 +
<br /><br />
 +
 
 +
<figure>
 +
<img class="displayed" src="https://static.igem.org/mediawiki/2015/7/7b/Lanes_1-14.png">
 +
<figcaption>Picture K: Land 1: DNA Ladder Marker,<br />
 +
Lane 2-7: (J61100+R0062)+EcoRI+PstI,<br />
 +
Lane 8: J61100+EcoRI+XbaI,<br />
 +
Lane 9-14: (J61100+R0062)+XbaI</figcaption>
 +
</figure>
 +
<br /><br />
 +
 
 +
<figure>
 +
<img class="displayed" src="https://static.igem.org/mediawiki/2015/8/8e/Lanes_1-5.png">
 +
<figcaption>Picture L: Lane 1: DNA Ladder Marker,<br />
 +
Lane 2-4: (R0062+J61100+I732006)+XbaI+PstI,<br />
 +
Lane 5: I732006+XbaI+PstI</figcaption>
 +
</figure>
 +
<br /><br />
 +
 
 +
<figure>
 +
<img class="displayed" src="https://static.igem.org/mediawiki/2015/f/fe/Lanes_1-8-2.png">
 +
<figcaption>Picture M: Lane 1 : DNA Ladder marker,<br />
 +
Lane 2 : J61100+EcoRI,<br />
 +
Lane 3 : (R0062+J61100)+EcoRI,<br />
 +
Lane 4 : (R0062+J61100+I732006)+EcoRI,<br />
 +
Lane 5 : (K823017+ R0062+J61100+I732006)+EcoRI,<br />
 +
Lane 6 : (R0062+J61100)+EcoRI+PstI,<br />
 +
Lane 7 : (R0062+J61100+I732006)+EcoRI+PstI,<br />
 +
Lane 8 : (K823017+R0062+J61100+I732006)+EcoRI+PstI</figcaption>
 +
</figure>
 +
<br /><br />
 +
 
 +
�<br><br>
 +
<P style="text-align:left;">
 +
    <font color="black">
 +
After the construction of the plasmids, we ran some experiments with our model to make sure it worked.<br><br>
 +
</font></p>
 +
 
 +
<figure>
 +
<img src="https://static.igem.org/mediawiki/2015/9/99/Results-evkr.png">
 +
<figcaption>Picture N: Final Results</figcaption>
 +
</figure>
 +
<br /><br />
 +
 
 +
<P style="text-align:left;">
 +
    <font color="black">
 +
We confirmed that GFP was exhibited in reporter cells to which AHL was added.<br><br>
 +
 
 +
We confirmed that if the test cell was added to the GFP beforehand, GFP was not exhibited.<br><br>
  
 
                     </font>
 
                     </font>
                     </p>
+
                     </p>
 
</div>
 
</div>
 +
<br><br>
 +
 +
<p style="text-align:center;">
 +
<video controls>
 +
<source src="https://static.igem.org/mediawiki/2015/8/89/PlasmidConstructionProcess.mp4" type="video/mp4">
 +
</video></p>
 +
<br /><br />
  
<a href="#top" rel="" id="top" class="anchorLink"><img class="displayed" src="https://static.igem.org/mediawiki/2015/5/5b/Scroll_arrow_top_Black.png"></a>
+
<a href="#top" rel="" id="top" class="anchorLink"><img class="displayed" src="https://static.igem.org/mediawiki/2015/5/5b/Scroll_arrow_top_Black.png"></a>
 
  <h6 style="text-align:center;"> <font color="black">
 
  <h6 style="text-align:center;"> <font color="black">
 
                   To The Top
 
                   To The Top
 
               </font> </h6>
 
               </font> </h6>
+
 
     
+
 
 
         </section>
 
         </section>
  

Latest revision as of 17:10, 18 September 2015








PROJECT
-Result-



Result


Our project had two main stages: in the first stage, we made the relevant plasmids, and in the second stage, we tested the cells transformed with the resulting plasmids. On the second stage, we used control experiments to make sure that the results we got was our actual results. For the control tests, we used untransformed E. coli and pure AHL as control groups.

Through the experiments, we confirmed the successful recombination of the test plasmid and the reporter plasmid, and the successful transformation of the test cell and the reporter cell. To make sure everything goes as planned, we tested the plasmids not only at the end, but also after the addition of each part.

For our project, we engineered two main plasmids: the reporter plasmid and the test plasmid. The reporter cell, which are competent E. coli transformed by reporter plasmids, produces LuxR, and has an inducible promoter that is activated by a LuxR-AHL complex. This promoter activates the transcription of GFP, which is easily detectable. The test cell, which are competent E. coli transformed by test plasmids, produces lactonase, which breaks down AHL. For the test plasmid, we tried two different types of enzymes: AiiA, and LacZ or beta-galactosidase.

When the test cell is treated with AHL, it breaks down AHL. When the reporter cell is added to the mix, no GFP is produced, as there is no AHL, whereas GFP is produced when a control group is used instead of a test cell.

We used the plasmids J61100, C0060, C0062, K823017, J37032, R0062, I732006 We transformed and raised the cells in solid plates to observe growth.

Picture A: C0062


Picture B: C0060


Picture C: I732006


Picture D: R0062


Picture E: J37032


Picture F: K823017


We went through miniprep, then electrophoresis to confirm that the plasmids were the ones we wanted.

For our initial electrophoresis results for the individual plasmids, look at: Picture H, Picture I, Picture J, Picture K Lane 8, Picture L Lane 5, Picture M Lane 1.

After we confirmed that all the plasmids were as they should be, we started the recombination process.

We also checked on every step of the recombination process by electrophoresis.

Picture G: Ladder reference


Picture H: Lane 1 : DNA ladder marker,
Lane 3 : C0060+EcoRI+PstI,
Lane 6 : I732073+EcoRI+PstI,
Lane 7 : J37032+EcoRI+PstI,
Lane 8 : K823017+EcoRI+PstI


Picture I: Lane 1-3: C0062+EcoRI+PstI,
Lane 9: DNA Ladder Marker


Picture J: Lane 1: DNA Ladder Marker,
Lane 2-3: R0062+EcoRI+SpeI


Picture K: Land 1: DNA Ladder Marker,
Lane 2-7: (J61100+R0062)+EcoRI+PstI,
Lane 8: J61100+EcoRI+XbaI,
Lane 9-14: (J61100+R0062)+XbaI


Picture L: Lane 1: DNA Ladder Marker,
Lane 2-4: (R0062+J61100+I732006)+XbaI+PstI,
Lane 5: I732006+XbaI+PstI


Picture M: Lane 1 : DNA Ladder marker,
Lane 2 : J61100+EcoRI,
Lane 3 : (R0062+J61100)+EcoRI,
Lane 4 : (R0062+J61100+I732006)+EcoRI,
Lane 5 : (K823017+ R0062+J61100+I732006)+EcoRI,
Lane 6 : (R0062+J61100)+EcoRI+PstI,
Lane 7 : (R0062+J61100+I732006)+EcoRI+PstI,
Lane 8 : (K823017+R0062+J61100+I732006)+EcoRI+PstI




After the construction of the plasmids, we ran some experiments with our model to make sure it worked.

Picture N: Final Results


We confirmed that GFP was exhibited in reporter cells to which AHL was added.

We confirmed that if the test cell was added to the GFP beforehand, GFP was not exhibited.





To The Top