Difference between revisions of "Team:Cardiff Wales/Results"

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<img width=100% src="https://static.igem.org/mediawiki/2016/d/d1/T--Cardiff_Wales--Achievements.png"/>
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<h1>Results</h1>
 
<h1>Results</h1>
 
<br>
 
<br>
−
<br>
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−
<br>
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<h1>Projects</h1>
 
<h1>Projects</h1>
 
<hr>  
 
<hr>  
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<ul>
 
<ul>
 
   <li>Use of vector (pET16B) with non-expected insert doomed 1/3 of our potential constructs</li>
 
   <li>Use of vector (pET16B) with non-expected insert doomed 1/3 of our potential constructs</li>
−
   <li>Failure to clone Cas-LucC/N & Cas-LacZA/O into E.Coli</li>
+
   <li>Failure to clone Cas-LucC/N & into E.Coli</li>
−
   <li><b>As a result we have been unable to study the viablity of our proposed diagnostic tool</b></li>
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   <li>As a result we have been unable to study the viablity of our proposed diagnostic tool</li>
  
 
</ul>
 
</ul>
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  <div style="float:left; margin:0; width:50%;">
 
  <div style="float:left; margin:0; width:50%;">
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<h2>FUEL</h2>
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<h2>FUEL Project</h2>
 
<hr>
 
<hr>
 
<h3> Fuel Up </h3>
 
<h3> Fuel Up </h3>
 
  <ul>
 
  <ul>
−
   <li>We have successfully cloned our arabinose mkeima and Lux Operon-mKeima constructs</li>
+
   <li>We have successfully cloned and sequenced our arabinose-inducible pBAD:mKeima and pBAD:Lux Operon:mKeima constructs</li>
−
<li>We have demonstrated that the Lux Operon of the fusion construct is still functional</li>
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<li>We have demonstrated that the LuxOperon portion of LUXoperon:mKeima of the fusion construct is functional</li>
  
 
</ul>
 
</ul>
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<h3> Running on Empty </h3>
 
<h3> Running on Empty </h3>
 
<ul>
 
<ul>
−
<li>We have been unable to express mKeima from either of the constructs</li>
+
<li>We have been unable to show a red light output from the mKeima constructs </li>
−
<li>We have therefore been unable to observe any red shifting of the Lux Operon</li>
+
<li>We have therefore been unable to observe any red shifting of the light output generated by the Lux Operon</li>
−
<li><b>Again we have as a result of our issues with cloning, been unable to test our hypothesis.</b></li>
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</ul>
 
</ul>
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</div>
 
</div>
  
−
<p>There are allot of things that we have learnt from out first year and hopefully next year we will be in a much stronger position.
+
<p>The inaugural year of Cardiff iGEM has been an excellent learning experience. There is no doubt that next year we will be in a much stronger position to succeed in all aspects of the our project. We have already set the wheels in motion for a Cardiff_Wales iGEM 2017.  
−
If we were to do this experiment again then:
+
<br><br>
−
<li> We would likely use the pSB1C3 antibiotic resistance variants instead of the three distinct plasmids we chose to use (pSB1C3, pET16B, pCOLA-DUET). This would improve the control of copy number and therefore help uniform the expression of our constructs. P.S. it removes the chance of a mystery insert in one our plasmids.</li>
+
<p> <i>Therefore what did we specifically learn from this years iGEM project?</i>
−
<li> We really struggled with our cloning with many of our constructs failing to transform. We thought it may potentially be due to the size's of our constructs, so next time we'd look at potentially using electrocompetent cells.</li>
+
<li> We would likely use the different pSB1C3 antibiotic resistance variants instead of the three distinct plasmids we chose to use (pSB1C3, pET16B, pCOLA-DUET). This would improve the control of copy number and help maintain uniform expression of our constructs. We were unaware of the different pSB1C3 variants due to our focus on cloning biobricks into the registry-compliant chloramphenicol resistant pSB1C3.</li>
−
<li> A number of our primers did not behave as expected and meant often we couldn't inspect our transformed cultures without directly sequencing them. Next time we will set more time and resources to checking all our primers work appropriately.</li>
+
<li> A number of our primers did not work as predicted and meant that we often couldn't inspect our transformed cultures without using direct sequencing. Next time we will set more time and resources to checking all our primers work appropriately, although within the time-scale of a ten-week project this can be challenging as by the time you realise there is a problem, the project has significantly progressed.</li>
−
<li> When we started lab work the whole team was involved. This has the effect of making the work a bit fractured and disjointed, not to mention complicating the management of samples. When we spoke to other teams it became apparent that it was the norm to have only a few members designated to lab duties. We therefore decided to reduce the number of us in the lab, and split off into two groups with each one focusing on a project each. This is something we look to do from the start next time.</li>
+
<li> When we started our lab work, the whole team was involved. This has the effect of making the work a bit fractured and disjointed, not to mention complicating the management of samples. In discussion with other teams it became apparent that it was more routine to have only a few members designated to lab duties. We therefore decided to reduce the number of us in the lab, and split off into two groups with each one focusing on a project each. This is something we look to do from the start in 2017.</li>
−
<li> We didn't start our FUEL project until after half of our Lab time was up, this really restricted the amount of time we could devote to it. Next time we would aim to split into two lab groups that start at the same together but work independently, which again would improve the management of lab work. </li>
+
<li> We didn't start our FUEL project until after half of our lab time was up, this really restricted the amount of time we could devote to it. Next time we would aim to split into two lab groups that start at the same together but work independently, which again would improve the management of lab work. </li>
  
  
 
<div id=d>  
 
<div id=d>  
  
−
<h1>Human Practices</h1>
 
−
<hr>
 
 
<div style="float:left; margin:0; width:50%;">
 
<div style="float:left; margin:0; width:50%;">
−
<h2>Outreach</h2>
+
 
−
<hr>
+
 
−
<li>We engaged with the general public at numerous events</li>
+
 
−
<li>As part of this we participated in educational events for a range of ages</li>
+
−
<li>We have interacted with IGEM teams, businesses and the public via our social media pages</li>
+
−
<li>Succeeded in creating interactive 3D models for explaining synthetic biology</li>
+
  
 
</div>
 
</div>
 
<div style="float:left; margin:0; width:50%;">
 
<div style="float:left; margin:0; width:50%;">
−
<h2>Bioethics</h2>
 
−
<hr>
 
−
<li>We have explored the ethical implications of home testing</li>
 
−
<li>We have sought out the opinions and knowledge of a number of respected professionals in the field </li>
 
−
<li>We survey'd the public opinion on home testing kits and the ethical questions they pose</li>
 
  
  
−
</div>
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</div>
 
</div>
 
<div id=d>  
 
<div id=d>  
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<li> Provided <a href="https://2016.igem.org/Team:Oxford" target="_blank">Oxford </a> with FLIM analysis of their copper chelators.  </li>
 
<li> Provided <a href="https://2016.igem.org/Team:Oxford" target="_blank">Oxford </a> with FLIM analysis of their copper chelators.  </li>
−
<li> We have quantified ATP production from cells transformed with <a href="https://2016.igem.org/Team:WashU_StLouis" target="_blank">Washington's</a> phosphoenolpyruvate kinase and phosphoglycerate kinase constructs </li>
+
<li> We have quantified ATP production from cells transformed with <a href="https://2016.igem.org/Team:WashU_StLouis" target="_blank">Washington's</a> phosphoenolpyruvate kinase (PCK) and phosphoglycerate kinase (PGK) constructs </li>
 
<br>
 
<br>
−
<p>Unfortunately we were unable to practically collaborate with <a href="https://2016.igem.org/Team:Paris_Saclay" target="_blank">Sacley</a> due to failures in our related dCas9. <p>
+
<p>Unfortunately we were unable to practically collaborate with <a href="https://2016.igem.org/Team:Paris_Saclay" target="_blank">Saclay</a> due to failures in our related dCas9. In future years we aim to increase our number of collaborations. We dedicated most of our lab time to cloning our own constructs, which meant there wasn't much time, or manpower left for collaborations. </p>
−
<p>In future years we aim to increase our number of collaborations. We dedicated most of our lab time to cloning our own constructs, which meant there wasn't much time, or manpower left for collaborations. </p>
+
 
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Latest revision as of 03:24, 3 December 2016

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Results


Projects

Cas-Find

Cas-Found

  • IPTG inducible CRISPR-Cas9 guide RNA targeted to rRNA Reverse
  • Investigated viable systems for future investigation

Can't-Find

  • Use of vector (pET16B) with non-expected insert doomed 1/3 of our potential constructs
  • Failure to clone Cas-LucC/N & into E.Coli
  • As a result we have been unable to study the viablity of our proposed diagnostic tool

FUEL Project

Fuel Up

  • We have successfully cloned and sequenced our arabinose-inducible pBAD:mKeima and pBAD:Lux Operon:mKeima constructs
  • We have demonstrated that the LuxOperon portion of LUXoperon:mKeima of the fusion construct is functional

Running on Empty

  • We have been unable to show a red light output from the mKeima constructs
  • We have therefore been unable to observe any red shifting of the light output generated by the Lux Operon

The inaugural year of Cardiff iGEM has been an excellent learning experience. There is no doubt that next year we will be in a much stronger position to succeed in all aspects of the our project. We have already set the wheels in motion for a Cardiff_Wales iGEM 2017.

Therefore what did we specifically learn from this years iGEM project?

  • We would likely use the different pSB1C3 antibiotic resistance variants instead of the three distinct plasmids we chose to use (pSB1C3, pET16B, pCOLA-DUET). This would improve the control of copy number and help maintain uniform expression of our constructs. We were unaware of the different pSB1C3 variants due to our focus on cloning biobricks into the registry-compliant chloramphenicol resistant pSB1C3.
  • A number of our primers did not work as predicted and meant that we often couldn't inspect our transformed cultures without using direct sequencing. Next time we will set more time and resources to checking all our primers work appropriately, although within the time-scale of a ten-week project this can be challenging as by the time you realise there is a problem, the project has significantly progressed.
  • When we started our lab work, the whole team was involved. This has the effect of making the work a bit fractured and disjointed, not to mention complicating the management of samples. In discussion with other teams it became apparent that it was more routine to have only a few members designated to lab duties. We therefore decided to reduce the number of us in the lab, and split off into two groups with each one focusing on a project each. This is something we look to do from the start in 2017.
  • We didn't start our FUEL project until after half of our lab time was up, this really restricted the amount of time we could devote to it. Next time we would aim to split into two lab groups that start at the same together but work independently, which again would improve the management of lab work.

    • Collaborations

  • Provided Oxford with FLIM analysis of their copper chelators.
  • We have quantified ATP production from cells transformed with Washington's phosphoenolpyruvate kinase (PCK) and phosphoglycerate kinase (PGK) constructs

    • Unfortunately we were unable to practically collaborate with Saclay due to failures in our related dCas9. In future years we aim to increase our number of collaborations. We dedicated most of our lab time to cloning our own constructs, which meant there wasn't much time, or manpower left for collaborations.