Team:Lethbridge/Attributions

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=Collaboration - Cell Viability in Tailings Pond Water=
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We were asked by the Calgary iGEM team to to test the viability of their chassis to survive in tailings pond water. If <i>E. coli</i> DH5α cells containing one of their constructs can grow normally in medium prepared with tailings water, we can be confident that the chassis can be applied to tailings pond water in general.
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==Materials and Methods==
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50 mL of LB medium was made with tailings water and filtered through MILLEX filter Unit GS MF-Millipore MCE Membrane 0.22 µm in order to sterilize it and appropriate antibiotics were added. The tailings LB was inoculated with <i>E. coli</i> DH5α cells containing BBa_K331009 to an OD<sub>600</sub> of 0.1. The flask was incubated at 37˚C with shaking for 7 hours, during which photometric readings were taken every 30. Photometric readings were taken using the Pharmacia Biotech Ultrospec 3000, as the OD<sub>600</sub> readings reached 1.0 the solutions was diluted with LB media to keep the OD<sub>600</sub> reading between 0.1 and 1.0. The same protocol was used to observe cell growth of <i>E. coli</i> DH5α cells containing BBa_K331009 in 50 mL of LB medium made with MilliQ H<sub>2</sub>O.
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==Results==
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[[image:uofltailings.png|center|400px]]
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<b>Figure 1.</b>    OD<sub>600</sub> of <i>E. coli</i> DH5α cells containing BBa_K331009 cultured in LB medium made with tailings water (red) and LB medium made with MilliQ H<sub>2</sub>O (black).  Performed by our team.
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[[image:uofltailings2.png|center|400px]]
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<b>Figure 2.</b>    OD<sub>600</sub> of <i>E. coli</i> DH5α cells containing BBa_K331009 cultured in LB medium made with tailings water (red and blue) and LB medium made with MilliQ H<sub>2</sub>O (black).  Performed independently by the Calgary 2011 Team.
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==Conclusion==
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As seen in Figure 1, <i>E. coli</i> DH5α cells were able to grow in LB medium made with tailings water, suggesting that the chassis will survive for use in tailings pond water.
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=<font color="black">Attributions & Contributions=
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The University of Lethbridge iGEM team would like to acknowledge the following people for their attributions and expertise which added to the success of our project, it is greatly appreciated, and has helped us gain new knowledge and skills.
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<a href="https://2011.igem.org/Team:Lethbridge">
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===Dr. A. William Smith===
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<img src="https://static.igem.org/mediawiki/2010/2/22/UofLHome.jpg" width="80"/>
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The Department of New Media at the University of Lethbridge – For his assistance, knowledge, leadership and hard work, in the Biospirits film project.
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===Mr. Doug Bray===
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The Faculty of Arts and Science at the University of Lethbridge –  For his training and guidance of team members in the use of transmission electron microscopy and scanning electron microscopy.
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===Dr. Marc Roussel===
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The Department of Chemistry and Biochemistry at the University of Lethbridge – For his assistance with protein modeling.
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<th><a href="https://2011.igem.org/Team:Lethbridge/Project">
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===Mr. David Lain Huston and the Arnott Labs===
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<img src="https://static.igem.org/mediawiki/2010/8/8d/UofLProjectbutton.jpg" width="80"/>
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The Institute of Cellular, Molecular, and Systems biology in Glasgow – For sending the team genes from the Xylene degradation pathway.
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===Others involved in bioSpirits film===
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Members of the academic instructional community:
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* Dennis Connolly [Math]
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* James Graham [New Media]
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Students:
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* Jeff Henry
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* Zsuzsi Papp
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* Jane Collin
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* J.P. Pehl
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* Colin McKague.
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Community members:
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* Two popular rock radio personalities: Vince and Rosie. [They interviewed Smith for their show.]
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* Justin Alho [Elementary]
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* Ryan Alho and Shayna FullerSmith [Middleschool]
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* Sharra FullerSmith [Highschool]
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* Christina Zehn
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* Neal Alho
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* Dan Lewis
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* Ruth Connolly
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* Patricia MacNally
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<th><a href="https://2011.igem.org/Team:Lethbridge/Safety">
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===Wet Lab Work===
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We are proud to say that all wet lab work was accomplished by the undergraduate members of our iGEM team.
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==<font color="white">Attributions & Contributions==
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<font color="white">Each team must clearly attribute work done by the team on this page.  They must distinguish work done by the team from work done by others, including the host labs, advisors, instructors, graduate students, and postgraduate masters students.
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Latest revision as of 00:47, 29 October 2011





Contents

Collaboration - Cell Viability in Tailings Pond Water

We were asked by the Calgary iGEM team to to test the viability of their chassis to survive in tailings pond water. If E. coli DH5α cells containing one of their constructs can grow normally in medium prepared with tailings water, we can be confident that the chassis can be applied to tailings pond water in general.

Materials and Methods

50 mL of LB medium was made with tailings water and filtered through MILLEX filter Unit GS MF-Millipore MCE Membrane 0.22 µm in order to sterilize it and appropriate antibiotics were added. The tailings LB was inoculated with E. coli DH5α cells containing BBa_K331009 to an OD600 of 0.1. The flask was incubated at 37˚C with shaking for 7 hours, during which photometric readings were taken every 30. Photometric readings were taken using the Pharmacia Biotech Ultrospec 3000, as the OD600 readings reached 1.0 the solutions was diluted with LB media to keep the OD600 reading between 0.1 and 1.0. The same protocol was used to observe cell growth of E. coli DH5α cells containing BBa_K331009 in 50 mL of LB medium made with MilliQ H2O.

Results

Uofltailings.png

Figure 1. OD600 of E. coli DH5α cells containing BBa_K331009 cultured in LB medium made with tailings water (red) and LB medium made with MilliQ H2O (black). Performed by our team.

Uofltailings2.png

Figure 2. OD600 of E. coli DH5α cells containing BBa_K331009 cultured in LB medium made with tailings water (red and blue) and LB medium made with MilliQ H2O (black). Performed independently by the Calgary 2011 Team.

Conclusion

As seen in Figure 1, E. coli DH5α cells were able to grow in LB medium made with tailings water, suggesting that the chassis will survive for use in tailings pond water.


Attributions & Contributions

The University of Lethbridge iGEM team would like to acknowledge the following people for their attributions and expertise which added to the success of our project, it is greatly appreciated, and has helped us gain new knowledge and skills.

Dr. A. William Smith

The Department of New Media at the University of Lethbridge – For his assistance, knowledge, leadership and hard work, in the Biospirits film project.

Mr. Doug Bray

The Faculty of Arts and Science at the University of Lethbridge – For his training and guidance of team members in the use of transmission electron microscopy and scanning electron microscopy.

Dr. Marc Roussel

The Department of Chemistry and Biochemistry at the University of Lethbridge – For his assistance with protein modeling.

Mr. David Lain Huston and the Arnott Labs

The Institute of Cellular, Molecular, and Systems biology in Glasgow – For sending the team genes from the Xylene degradation pathway.

Others involved in bioSpirits film

Members of the academic instructional community:

  • Dennis Connolly [Math]
  • James Graham [New Media]

Students:

  • Jeff Henry
  • Zsuzsi Papp
  • Jane Collin
  • J.P. Pehl
  • Colin McKague.

Community members:

  • Two popular rock radio personalities: Vince and Rosie. [They interviewed Smith for their show.]
  • Justin Alho [Elementary]
  • Ryan Alho and Shayna FullerSmith [Middleschool]
  • Sharra FullerSmith [Highschool]
  • Christina Zehn
  • Neal Alho
  • Dan Lewis
  • Ruth Connolly
  • Patricia MacNally

Wet Lab Work

We are proud to say that all wet lab work was accomplished by the undergraduate members of our iGEM team.