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

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<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>
 
<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>
 
<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 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> 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> 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 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> 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>

Revision as of 11:35, 18 October 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 & Cas-LacZA/O into E.Coli
  • As a result we have been unable to study the viablity of our proposed diagnostic tool

FUEL

Fuel Up

  • We have successfully cloned our arabinose mkeima and Lux Operon-mKeima constructs
  • We have demonstrated that the Lux Operon of the fusion construct is still functional

Running on Empty

  • We have been unable to express mKeima from either of the constructs
  • We have therefore been unable to observe any red shifting of the Lux Operon
  • Again we have as a result of our issues with cloning, been unable to test our hypothesis.

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. If we were to do this experiment again then:

  • 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.
  • 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.
  • 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.
  • 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.
  • 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.

    • Human Practices

    Outreach

  • We engaged with the general public at numerous events
  • As part of this we participated in educational events for a range of ages
  • We have interacted with IGEM teams, businesses and the public via our social media pages
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    • Bioethics

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    • Collaborations

    Here you can describe the results of your project and your future plans.

    What should this page contain?
    • Clearly and objectively describe the results of your work.
    • Future plans for the project
    • Considerations for replicating the experiments
    Project Achievements

    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.

    • A list of linked bullet points of the successful results during your project
    • 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.
    Inspiration

    See how other teams presented their results.