Difference between revisions of "Team:Edinburgh/Characterisation Part"
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<li><a href="https://2015.igem.org/Team:Edinburgh/DNPBiosensor">DNP Biosensor</a></li> | <li><a href="https://2015.igem.org/Team:Edinburgh/DNPBiosensor">DNP Biosensor</a></li> | ||
<li><a href="https://2015.igem.org/Team:Edinburgh/PMABiosensor">PMA Biosensor</a></li> | <li><a href="https://2015.igem.org/Team:Edinburgh/PMABiosensor">PMA Biosensor</a></li> | ||
| − | <li><a href="https://2015.igem.org/Team:Edinburgh/CBD">Making it Stick</a></li> | + | <li><a href="https://2015.igem.org/Team:Edinburgh/CBD">Making it Stick</a></li> |
| − | <li><a href="https://2015.igem.org/Team:Edinburgh/Results"> | + | <li><a href="https://2015.igem.org/Team:Edinburgh/Results">Limits of Detection</a></li> |
</ul> | </ul> | ||
</li> | </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> | ||
<ul class="dropdown-menu" role="menu"> | <ul class="dropdown-menu" role="menu"> | ||
| − | + | <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> | ||
| − | + | <li><a href="https://2015.igem.org/Team:Edinburgh/Part_Collection">Part Collection</a> </li> | |
<li><a href="https://2015.igem.org/Team:Edinburgh/Improved_Part">Improved Parts</a></li> | <li><a href="https://2015.igem.org/Team:Edinburgh/Improved_Part">Improved Parts</a></li> | ||
<li><a href="https://2015.igem.org/Team:Edinburgh/Characterisation_Part">Improved Characterisation</a></li> | <li><a href="https://2015.igem.org/Team:Edinburgh/Characterisation_Part">Improved Characterisation</a></li> | ||
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</ul> | </ul> | ||
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| − | <li><a href="https://2015.igem.org/Team:Edinburgh/MedalCriteria"> | + | <li><a href="https://2015.igem.org/Team:Edinburgh/MedalCriteria">Accomplishments</a></li> |
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<p class="intro-text"> | <p class="intro-text"> | ||
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| − | <a href="#cd-team"> | + | <div align="center"> |
| − | + | <a href="#cd-team"> | |
| − | + | <span class="arrowtext">Scroll down to read more</span> | |
| − | + | <img src="https://static.igem.org/mediawiki/2014/3/3e/Aalto_Helsinki_Nuoli.png" class="arrow"> | |
| + | </a> | ||
| + | </div> | ||
</div> | </div> | ||
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<h3 class="section-subheading text-muted"> | <h3 class="section-subheading text-muted"> | ||
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| + | We improved and expanded characterisation on the part BBa_K1321357, a carbohydrate binding domain (CBD) submitted by the Imperial College 2014 team. After confirming expression through SDS PAGE we incubated iso-standard paper chads from Whatman 54 filter paper in a crude cell lysate containing the CBD bound to super folded GFP for ten minutes. We then let the protein saturated chads sit in a large volume of 1 x PBS buffer at pH 7.4 (to remain consistent with imperial’s initial characterisation.) At different time points chads were taken from the buffer and placed into a 96 well plate, over 30 minutes. The decrease in fluorescence over time was indicative of relative binding affinity, to Whatman 54 paper. The graph below shows our results. | ||
| + | </p> | ||
| + | <br> | ||
| + | <br> | ||
| + | <div align="center"> | ||
| + | <img src="https://static.igem.org/mediawiki/2015/e/ec/Dissociation_curve_of_sfGFP-CBDcex.jpeg" class="img-responsive"> | ||
| + | </div> | ||
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| + | <br> | ||
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| + | We improved and expanded characterisation on the part BBa_K1321356, a carbohydrate binding domain (CBD) submitted by the Imperial College 2014 team. After confirming expression through SDS PAGE we incubated iso-standard paper chads from Whatman 54 filter paper in a crude cell lysate containing the CBD bound to super folded GFP for ten minutes. We then let the protein saturated chads sit in a large volume of 1 x PBS buffer at pH 7.4 (to remain consistent with imperial’s initial characterisation.) At different time points 5 chads were taken from the buffer and placed into a 96 well plate, over 30 minutes. The decrease in fluorescence over time was indicative of relative binding affinity, to Whatman 54 paper. The graph below shows our results. | ||
| + | </p> | ||
| + | <br> | ||
| + | <br> | ||
| + | <div align="center"> | ||
| + | <img src="https://static.igem.org/mediawiki/2015/5/50/5.20_K.jpeg" class="img-responsive"> | ||
| + | </div> | ||
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| + | </p> | ||
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</h3> | </h3> | ||
</div> | </div> | ||
</div> | </div> | ||
| + | </div> | ||
| + | </section> | ||
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Latest revision as of 18:56, 20 November 2015
We improved and expanded characterisation on the part BBa_K1321357, a carbohydrate binding domain (CBD) submitted by the Imperial College 2014 team. After confirming expression through SDS PAGE we incubated iso-standard paper chads from Whatman 54 filter paper in a crude cell lysate containing the CBD bound to super folded GFP for ten minutes. We then let the protein saturated chads sit in a large volume of 1 x PBS buffer at pH 7.4 (to remain consistent with imperial’s initial characterisation.) At different time points chads were taken from the buffer and placed into a 96 well plate, over 30 minutes. The decrease in fluorescence over time was indicative of relative binding affinity, to Whatman 54 paper. The graph below shows our results.
We improved and expanded characterisation on the part BBa_K1321356, a carbohydrate binding domain (CBD) submitted by the Imperial College 2014 team. After confirming expression through SDS PAGE we incubated iso-standard paper chads from Whatman 54 filter paper in a crude cell lysate containing the CBD bound to super folded GFP for ten minutes. We then let the protein saturated chads sit in a large volume of 1 x PBS buffer at pH 7.4 (to remain consistent with imperial’s initial characterisation.) At different time points 5 chads were taken from the buffer and placed into a 96 well plate, over 30 minutes. The decrease in fluorescence over time was indicative of relative binding affinity, to Whatman 54 paper. The graph below shows our results.
