Difference between revisions of "Team:Edinburgh/Composite Part"

 
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                   <a href="https://2015.igem.org/Team:Edinburgh">Home</a></li>
 
                   <a href="https://2015.igem.org/Team:Edinburgh">Home</a></li>
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                    <a href="#" class="dropdown-toggle" data-toggle="dropdown" role="button" aria-expanded="false">About<span class="caret"></span></a>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Team">Team</a></li>
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                       <li><a href="https://2015.igem.org/Team:Edinburgh/Description">Description</a></li>
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                       <li><a href="https://2015.igem.org/Team:Edinburgh/Description">Overview</a></li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Experiments">Experiments</a> </li>
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                    <!-- <li><a href="https://2015.igem.org/Team:Edinburgh/Experiments">Experiments</a> </li> -->
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                       <li><a href="https://2015.igem.org/Team:Edinburgh/Project/Protocols">Protocols</a> </li>
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                       <li><a href="https://2015.igem.org/Team:Edinburgh/HeroinBiosensor">Heroin Biosensor</a></li>
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                       <li><a href="https://2015.igem.org/Team:Edinburgh/Results">Results</a></li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/DNPBiosensor">DNP Biosensor</a></li>
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                       <li><a href="https://2015.igem.org/Team:Edinburgh/Design">Design</a></li>
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                       <li><a href="https://2015.igem.org/Team:Edinburgh/PMABiosensor">PMA Biosensor</a></li>
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                       <li><a href="https://2015.igem.org/Team:Edinburgh/CBD">Making it Stick</a></li>           
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Results">Limits of Detection</a></li>
 
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                     <a href="#" class="dropdown-toggle" data-toggle="dropdown" role="button" aria-expanded="false">Parts<span class="caret"></span></a>
 
                     <a href="#" class="dropdown-toggle" data-toggle="dropdown" role="button" aria-expanded="false">Parts<span class="caret"></span></a>
 
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                       <li><a href="https://2015.igem.org/Team:Edinburgh/Parts">Team Parts</a></li>
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                       <li><a href="https://2015.igem.org/Team:Edinburgh/Parts">Team Parts</a></li>  
 
                       <li><a href="https://2015.igem.org/Team:Edinburgh/Basic_Part">Basic Parts</a></li>
 
                       <li><a href="https://2015.igem.org/Team:Edinburgh/Basic_Part">Basic Parts</a></li>
 
                       <li><a href="https://2015.igem.org/Team:Edinburgh/Composite_Part">Composite Parts</a></li>
 
                       <li><a href="https://2015.igem.org/Team:Edinburgh/Composite_Part">Composite Parts</a></li>
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                       <li><a href="https://2015.igem.org/Team:Edinburgh/Part_Collection">Part Collection</a> </li>
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                       <li><a href="https://2015.igem.org/Team:Edinburgh/Part_Collection">Part Collection</a> </li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Improved_Part">Improved Parts</a></li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Characterisation_Part">Improved Characterisation</a></li>
 
                     </ul>
 
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                  <li><a href="https://2015.igem.org/Team:Edinburgh/Attributions">Attributions</a></li>
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                  <li><a href="https://2015.igem.org/Team:Edinburgh/Practices">Policy and Practices</a></li>
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                    <a href="#" class="dropdown-toggle" data-toggle="dropdown" role="button" aria-expanded="false">Dry Lab<span class="caret"></span></a>
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                  <li><a href="https://2015.igem.org/Team:Edinburgh/Modeling">Modeling</a></li>
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                   <li><a href="https://2015.igem.org/Team:Edinburgh/InterLab">InterLab</a></li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Software">Software</a></li>
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                   <li><a href="entrepreneurship.html">Entrepreneurship</a></li>
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                    </ul>
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                  <li><a href="collaborations.html">Collaborations</a></li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Practices">Our Story</a></li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Safety">Safety</a> </li>
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                    <a href="#" class="dropdown-toggle" data-toggle="dropdown" role="button" aria-expanded="false">Notebook<span class="caret"></span></a>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Project/Protocols">Protocols</a></li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Notebook/HeroinPurity">Heroin Purity</a></li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Notebook/PMADetection">PMA Detection</a> </li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Notebook/DNPDetection">DNP Detection</a> </li>
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                      <li><a href="https://2015.igem.org/Team:Edinburgh/Notebook/FluidDynamics">Fluid Dynamics</a> </li>
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                    </ul>
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                  </li>
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                  <li><a href="https://2015.igem.org/Team:Edinburgh/MedalCriteria">Accomplishments</a></li> 
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            </ul>
 
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                 <h1 class="brand-heading">Composite Parts</h1>
 
                 <h1 class="brand-heading">Composite Parts</h1>
 
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                    <a href="#cd-team">
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                        <span class="arrowtext">Scroll down to read more</span>
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                        <img src="https://static.igem.org/mediawiki/2014/3/3e/Aalto_Helsinki_Nuoli.png" class="arrow">
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                     <h2 class="section-heading">Foreword</h2>                     
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                     <h2 class="section-heading">mRFP</h2>                     
 
                 </div>
 
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                     <a data-toggle="collapse" data-parent="#accordion" href="#collapseOne">1. What is HTML?</a>
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                     <a data-toggle="collapse" data-parent="#accordion" href="#collapseOne">mRFP driven by LacI promoter (BBa_K1615090)</a>
  
 
                 </h4>
 
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                 <div class="panel-body">
 
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                     <p>HTML stands for HyperText Markup Language. HTML is the main markup language for describing the structure of Web pages. <a href="http://www.tutorialrepublic.com/html-tutorial/" target="_blank">Learn more.</a></p>
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                     <p>mRFP (BBa_K1615089) driven by the IPTG inducible PLlac 0-1 promoter (BBa_K314103). This allows the <i>mRFP</i> gene to express mRFP when induced with IPTG.
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</p>
  
 
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                     <a data-toggle="collapse" data-parent="#accordion" href="#collapseTwo">2. What is Bootstrap?</a>
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                     <a data-toggle="collapse" data-parent="#accordion" href="#collapseTwo">CBDcenA+Linker fused to mRFP in RFC25 (BBa_K1615091)</a>
  
 
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                     <p>Bootstrap is a powerful front-end framework for faster and easier web development. It is a collection of CSS and HTML conventions. <a href="http://www.tutorialrepublic.com/twitter-bootstrap-tutorial/" target="_blank">Learn more.</a></p>
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                     <p>mRFP (BBa_K1615089) fused to the C terminal of CBDcenA + C-terminal linker (BBa_K1321339) in RFC25. If folded properly, this allows mRFP to fluoresce while immobilised on cellulose.
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<br>
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<img src="https://static.igem.org/mediawiki/2015/2/20/CBD_RFP_folding.jpeg">
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<br>
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To test the RFP folding when fused to a CBD we placed 200µL of crude cell lysate in a 96 well plate and observed the fluorescence under a blue light transilluminator.
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                     <a data-toggle="collapse" data-parent="#accordion" href="#collapseThree">3. What is CSS?</a>
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                     <a data-toggle="collapse" data-parent="#accordion" href="#collapseFour">mRFP fused to CBDcenA+Linker in RFC25 (BBa_K16150092)</a>
  
 
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                     <p>CSS stands for Cascading Style Sheet. CSS allows you to specify various style properties for a given HTML element such as colors, backgrounds, fonts etc. <a href="http://www.tutorialrepublic.com/css-tutorial/" target="_blank">Learn more.</a></p>
+
                     <p>mRFP (BBa_K1615089) fused to the N terminal of CBDcenA + C terminal linker (BBa_K1321339) in RFC25. If folded properly, this allows mRFP to fluoresce while immobilised on cellulose.
 +
<br>
 +
<br>
 +
<img src="https://static.igem.org/mediawiki/2015/2/20/CBD_RFP_folding.jpeg">
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<br>
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<br>
 +
We tested the dissociation of this part on Whatman 54 using isostandard chads. This involved incubating the chads in the crude cell lysate for 10 minutes and then washing them with PBS for various periods time to test the binding affinity of this CBD.
 +
<br>
 +
<br>
 +
We tested the dissociation of this part on Whatman 54 using isostandard chads. This involved incubating the chads in the crude cell lysate for 10 minutes and then washing them with PBS for various periods time to test the binding affinity of this CBD.
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</p>
  
 
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                <h4 class="panel-title">
  
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                    <a data-toggle="collapse" data-parent="#accordion" href="#collapseFive">Linker +dCBD fused to mRFP in RFC25 (BBa_K1615093)
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</a>
  
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      <footer>
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                </h4>
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        <p class="pull-right"><a href="#">Back to top</a></p>
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        <p>&copy; 2015 EdiGEM &middot; <a href="#">Privacy</a> &middot; <a href="#">Terms</a></p>
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                <div class="panel-body">
 +
 +
                    <p>mRFP (BBa_K1615089) fused to the C terminal of dCBD + N terminal linker (BBa_K1321340). If folded properly, this allows mRFP to fluoresce while immobilised on cellulose.
 +
<br>
 +
<br>
 +
<img src="https://static.igem.org/mediawiki/2015/2/20/CBD_RFP_folding.jpeg">
 +
<br>
 +
To test the RFP folding when fused to a CBD we placed 200µL of crude cell lysate in a 96 well plate and observed the fluorescence under a blue light transilluminator.
 +
</p>
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Latest revision as of 19:10, 20 November 2015

Composite Parts

mRFP

mRFP driven by LacI promoter (BBa_K1615090)

mRFP (BBa_K1615089) driven by the IPTG inducible PLlac 0-1 promoter (BBa_K314103). This allows the mRFP gene to express mRFP when induced with IPTG.

CBDcenA+Linker fused to mRFP in RFC25 (BBa_K1615091)

mRFP (BBa_K1615089) fused to the C terminal of CBDcenA + C-terminal linker (BBa_K1321339) in RFC25. If folded properly, this allows mRFP to fluoresce while immobilised on cellulose.

To test the RFP folding when fused to a CBD we placed 200µL of crude cell lysate in a 96 well plate and observed the fluorescence under a blue light transilluminator.

mRFP fused to CBDcenA+Linker in RFC25 (BBa_K16150092)

mRFP (BBa_K1615089) fused to the N terminal of CBDcenA + C terminal linker (BBa_K1321339) in RFC25. If folded properly, this allows mRFP to fluoresce while immobilised on cellulose.



We tested the dissociation of this part on Whatman 54 using isostandard chads. This involved incubating the chads in the crude cell lysate for 10 minutes and then washing them with PBS for various periods time to test the binding affinity of this CBD.

We tested the dissociation of this part on Whatman 54 using isostandard chads. This involved incubating the chads in the crude cell lysate for 10 minutes and then washing them with PBS for various periods time to test the binding affinity of this CBD.

Linker +dCBD fused to mRFP in RFC25 (BBa_K1615093)

mRFP (BBa_K1615089) fused to the C terminal of dCBD + N terminal linker (BBa_K1321340). If folded properly, this allows mRFP to fluoresce while immobilised on cellulose.


To test the RFP folding when fused to a CBD we placed 200µL of crude cell lysate in a 96 well plate and observed the fluorescence under a blue light transilluminator.