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- | <html xmlns="http://www.w3.org/1999/xhtml"> | + | <div style="background-color:#000000; color:white"> |
- | <head>
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- | <meta http-equiv="Content-Type" content="text/html; charset=utf-8" />
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- | <title>Untitled Document</title>
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- | <script src="http://people.uleth.ca/~david.franz/igem3/SpryAssets/SpryTabbedPanels.js" type="text/javascript"></script>
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- | </head>
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| + | <table border="0" width="100%" style="background-color:#000000"> |
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| + | <image src="https://static.igem.org/mediawiki/2010/2/29/UofLteamlogo.jpg" width="200px"/> |
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| + | </th> |
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| + | <th> |
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| + | <image src="https://static.igem.org/mediawiki/2010/4/4e/UofLwewantyou.jpg" height="300px"/> |
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| + | </th> |
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| + | <th> |
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| + | <image src="https://static.igem.org/mediawiki/2010/2/29/UofLteamlogo.jpg" width="200px"/> |
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| + | </th> |
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| + | </tr> |
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| + | </table> |
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| + | <br> |
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| + | <align="centre"> |
| + | <table border="0" width="100%" style="background-color:#000000"> |
| + | |
| + | <tr> |
| + | |
| + | <th> |
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| + | <a href="https://2010.igem.org/Team:Lethbridge"> |
| + | <img src="https://static.igem.org/mediawiki/2010/2/22/UofLHome.jpg" width="80"/> |
| + | </a> |
| + | |
| + | </th> |
| + | |
| + | <th><a href="https://2010.igem.org/Team:Lethbridge/Team"> |
| + | <img src="https://static.igem.org/mediawiki/2010/0/0d/UofLTeam.jpg" width="80"/> |
| + | </a> |
| + | </th> |
| + | |
| + | <th><a href="https://2010.igem.org/Team:Lethbridge/Project"> |
| + | <img src="https://static.igem.org/mediawiki/2010/8/8d/UofLProjectbutton.jpg" width="80"/> |
| + | </a> |
| + | </th> |
| + | |
| + | <th><a href="https://2010.igem.org/Team:Lethbridge/Notebook/Lab_Work"> |
| + | <img src="https://static.igem.org/mediawiki/2010/7/73/UofLNotebookbutton.jpg" width="80"/> |
| + | </a> |
| + | </th> |
| + | |
| + | <th><a href="https://2010.igem.org/Team:Lethbridge/Parts"> |
| + | <img src="https://static.igem.org/mediawiki/2010/8/84/UofLPartsSubmittedToTheRegistrybutton.jpg" width="80"/> |
| + | </a> |
| + | </th> |
| + | |
| + | <th><a href="https://2010.igem.org/Team:Lethbridge/Modeling"> |
| + | <img src="https://static.igem.org/mediawiki/2010/e/e1/UofLModelingbutton.jpg" width="80"/> |
| + | </a> |
| + | </th> |
| + | |
| + | <th><a href="https://2010.igem.org/Team:Lethbridge/Ethics"> |
| + | <img src="https://static.igem.org/mediawiki/2010/2/26/UofLEthicsbutton.jpg" width="80"/> |
| + | </a> |
| + | </th> |
| + | |
| + | <th><a href="https://2010.igem.org/Team:Lethbridge/Safety"> |
| + | <img src="https://static.igem.org/mediawiki/2010/0/00/UofLSafetybutton.jpg" width="80"/> |
| + | </a> |
| + | </th> |
| + | |
| + | <th><a href="https://2010.igem.org/Team:Lethbridge/Art"> |
| + | <img src="https://static.igem.org/mediawiki/2010/0/0a/UofLArt.jpg" width="80"/> |
| + | </a> |
| + | </th> |
| + | |
| + | <th><a href="https://2010.igem.org/Team:Lethbridge/News"> |
| + | <img src="https://static.igem.org/mediawiki/2010/c/c3/UofLNewsButton.jpg" width="80"/> |
| + | </a> |
| + | </th> |
| + | </table> |
| + | </body> |
| + | </html> |
| + | <hr> |
| + | <center> |
| + | Check out these important project links! |
| + | </center> |
| + | <html> |
| <body> | | <body> |
- | <p>University of Lethbridge IGEM team</p> | + | <center> |
- | <p> </p>
| + | <table border="0" width="20%" style="background-color:#000000"> |
- | <div id="TabbedPanels1" class="TabbedPanels"> | + | <tr> |
- | <ul class="TabbedPanelsTabGroup">
| + | |
- | <li class="TabbedPanelsTab" tabindex="0">Main</li>
| + | <th><a href="https://2010.igem.org/Team:Lethbridge/Judging"> |
- | <li class="TabbedPanelsTab" tabindex="0">Team</li>
| + | <img src="https://static.igem.org/mediawiki/2010/f/fd/UofLjudgingbutton.jpg" width="60"/> |
- | <li class="TabbedPanelsTab" tabindex="0">Projects</li>
| + | </a> |
- | <li class="TabbedPanelsTab" tabindex="0">Notebook</li>
| + | </th> |
- | <li class="TabbedPanelsTab" tabindex="0">Meetings</li>
| + | |
- | <li class="TabbedPanelsTab" tabindex="0">Parts</li>
| + | <th><a href="https://2010.igem.org/Team:Lethbridge/Results"> |
- | <li class="TabbedPanelsTab" tabindex="0">Collaborations</li>
| + | <img src="https://static.igem.org/mediawiki/2010/3/3f/UofLresultsbutton.jpg" width="60"/> |
- | </ul>
| + | </a> |
- | <div class="TabbedPanelsContentGroup">
| + | </th> |
- | <div class="TabbedPanelsContent">blah balh balh</div>
| + | |
- | <div class="TabbedPanelsContent">
| + | <th><a href="https://2010.igem.org/Team:Lethbridge/Collaboration"> |
- | <p>Our team is based yada yada</p>
| + | <img src="https://static.igem.org/mediawiki/2010/b/b2/UofLcollaborationbutton.png" width="60"/> |
- | <div id="TabbedPanels2" class="TabbedPanels">
| + | </a> |
- | <ul class="TabbedPanelsTabGroup">
| + | </th> |
- | <li class="TabbedPanelsTab" tabindex="0">Team Pics</li>
| + | |
- | <li class="TabbedPanelsTab" tabindex="0">Tab 3</li>
| + | |
- | <li class="TabbedPanelsTab" tabindex="0">Team bios</li>
| + | |
- | </ul>
| + | |
- | <div class="TabbedPanelsContentGroup">
| + | |
- | <div class="TabbedPanelsContent">Team pictures</div>
| + | |
- | <div class="TabbedPanelsContent">Content 3</div>
| + | |
- | <div class="TabbedPanelsContent">Pics of us</div>
| + | |
- | </div>
| + | |
- | </div>
| + | |
- | <p> </p>
| + | |
- | <p> </p>
| + | |
- | <p> </p>
| + | |
- | <p> </p>
| + | |
- | <p> </p>
| + | |
- | </div>
| + | |
- | <div class="TabbedPanelsContent">
| + | |
- | <p>Overview</p>
| + | |
- | <div id="TabbedPanels3" class="TabbedPanels">
| + | |
- | <ul class="TabbedPanelsTabGroup">
| + | |
- | <li class="TabbedPanelsTab" tabindex="0">Project 1</li>
| + | |
- | <li class="TabbedPanelsTab" tabindex="0">Porejct 2</li>
| + | |
- | </ul>
| + | |
- | <div class="TabbedPanelsContentGroup">
| + | |
- | <div class="TabbedPanelsContent">Content 1</div>
| + | |
- | <div class="TabbedPanelsContent">Content 2</div>
| + | |
- | </div>
| + | |
- | </div>
| + | |
- | <p> </p>
| + | |
- | <p> </p>
| + | |
- | <p> </p>
| + | |
- | </div>
| + | |
- | <div class="TabbedPanelsContent"><p> </p>
| + | |
- | <div id="TabbedPanels4" class="TabbedPanels">
| + | |
- | <ul class="TabbedPanelsTabGroup">
| + | |
- | <li class="TabbedPanelsTab" tabindex="0">Lab Work</li>
| + | |
- | <li class="TabbedPanelsTab" tabindex="0">Common Protocols</li>
| + | |
- | </ul>
| + | |
- | <div class="TabbedPanelsContentGroup">
| + | |
- | <div class=TabbedPanelsContent>
| + | |
| | | |
- | <b><font size=+1>Jump to Month:<br>
| + | <tr> |
- | <a href=#april>April</a><br>
| + | |
- | <a href=#may>May</a><br></b><br>
| + | |
- | <a name="april"></a></font>
| + | |
- | <b><font size=+2>April 13/2010</font> (In the Lab: JV, AS)</b><br>
| + | |
- | <b>Objective:</b> Test Restriction Endonucleases for Activity<br>
| + | |
- | <b>Relevant Information:</b><br>
| + | |
- | Endonucleases available
| + | |
- | <table><table border="3">
| + | |
- | <tr><td>Endonuclease</td><td>Optimal Buffer**</td><td>Other Buffers</td></tr> | + | |
- | <tr><td>EcoRV</td><td>None</td><td>2xT(100%); O,G(50-100%)</td></tr>
| + | |
- | <tr><td>EcoRI</td><td>Red</td><td>O(100%);R(100%)*;2xT(100%)</td></tr>
| + | |
- | <tr><td>BcuI/SpeI</td><td>Tango</td><td>B(50-100%);G(50-100%)</td></tr>
| + | |
- | <tr><td>XbaI</td><td>Tango</td><td>B,G,2xT(50-100%)</td></tr>
| + | |
- | <tr><td>PstI</td><td>Orange</td><td>R(100%); B,G,T,2xT(50-100%)</td></tr>
| + | |
- | <tr><td>DpnI</td><td>Tango</td><td>B,G(100%): O,R,2xT(50-100%)</td></tr>
| + | |
| </table> | | </table> |
- | *Star Activity<br>
| + | </center> |
- | **Optimal Buffer from Fermentas<br><br>
| + | </body> |
- | Use pUC19 plasmid as test, it has cut sites for EcoRI, PstI, XbaI (unsure about BcuI/SpeI, DpnI but will try anyways), and none for EcoRV <br>
| + | </html> |
- | <u>Red Buffer:</u> EcoRI, PstI, Control (No Enzyme)<br>
| + | <hr> |
- | <u>Tango Buffer:</u> BcuI/SpeI, XbaI, DpnI, Control (No Enzyme><br><br>
| + | |
- | <u>Methods:</u> | + | <BLOCKQUOTE> |
- | Set up Master Mixes:
| + | =<font color="white">Project Description= |
- | <table><table border="3"> | + | |
- | <tr><td><b>Red MM</b></td><td>per tube (µL)</td><td>Total (µL)</td></tr>
| + | The tailings ponds that result from the extraction of heavy crude oil and bitumen (used to make synthetic crude oil) from the oil sands have used up vast amounts of fresh water and contain substantial quantities of useable organic matter. While most of the bitumen is extracted from the oil sands, some is left behind and added to the tailings ponds. The residual hydrocarbon compounds can be potentially extracted from the ponds and utilized as another source of fuel. Consequently, cleaning the tailings ponds and increasing efficiency of extraction from the oil sands. |
- | <tr><td>MilliQ H<sub>2</sub>0</td><td>13.75</td><td>55</td></tr>
| + | <br> |
- | <tr><td>Red Buffer (10x)</td><td>2</td><td>7</td></tr>
| + | |
- | <tr><td>pUC19 (10pg/µL)</td><td>2</td><td>7</td></tr> | + | We wish to develop and characterize a BioBrick, that can breakdown some of the more prominent toxic organic compounds found in the tailing ponds to a more useable form. We are currently targeting catechol, a aromatic compound shown to be degraded by bacteria living in the tailings ponds (Kato <i>et al.</i>, 2001). Catechol is being converted into 2-hydroxymuconic semialdehyde, which we later hope to further convert into a useful hydrocarbon compound. |
- | <tr><td><b>Total</b></td><td>19.75</td><td>69</td></tr> | + | <br> |
- | </table><br> | + | |
- | <table><table border="3">
| + | Additionally, we plan to target our <html><a href="https://2010.igem.org/Team:Lethbridge/Project/Catechol_Degradation"><font color="#00DC00">catechol degrading enzyme</font></a></html> into a <html><a href="https://2010.igem.org/Team:Lethbridge/Project/Compartamentalization"><font color="#00DC00"> microcompartment</font></a></html> which the <html><a href="https://2009.igem.org/Team:Lethbridge" target="new"><font color="#00DC00"> Lethbridge 2009</font></a></html> team began the work on. By compartmentalizing the converted catechol, were trying to develop a way of easily removing the useful hydrocarbon product from the tailings ponds. As a proof of principle, we will target the catechol degradation enzyme into the negatively charged microcompartment by the use of a poly-arginine tag. Furthermore, to avoid adding a new species into the oil sands environment we plan on using the <html><a href="https://2010.igem.org/Team:Lethbridge/Project/DNA_Degradation"><font color="#00DC00"> DNA digestion part</font></a></html> created by <html><a href="https://2007.igem.org/Berkeley_UC" target="new"><font color="#00DC00"> Berkley in 2007</font></a></html> to render our <i>Escherichia coli </i>cells unable to reproduce or able to horizontally transfer its genes. |
- | <tr><td><b>Tango MM</b></td><td>per tube (µL)</td><td>Total (µL)</td></tr>
| + | <br> |
- | <tr><td>MilliQ H<sub>2</sub>0</td><td>13.75</td><td>55</td></tr>
| + | |
- | <tr><td>Tango Buffer (10x)</td><td>2</td><td>7</td></tr> | + | Finally, we will be continuing to explore the novel method of the mass production of <html><a href="https://2010.igem.org/Team:Lethbridge/Project/Magnetic_Nanoparticles"><font color="#00DC00"> uniform iron nanoparticles</font></a></html>, which is more efficient and cost effective than current methods (Prozorov <i>et al.</i>, 2007). To optimize the production of nanoparticles we are attaching signal peptide sequences to export the protein to different areas of the cell. By attaching these signal peptides and having the protein directed to certain areas of the cell we hope to find which area is most productive to produce nanoparticles. |
- | <tr><td>pUC19 (10pg/µL)</td><td>2</td><td>7</td></tr> | + | <br> |
- | <tr><td><b>Total</b></td><td>19.75</td><td>69</td></tr>
| + | |
- | </table><br>
| + | |
- | To each tube, add <b>19.75µL</b> of master mix and <b>0.25µL</b> of enzyme<br>
| + | Reference:<br> |
- | Incubated reaction mixes at 37<sup>o</sup>C (Start:7:00pm; End:7:45pm)<br>
| + | Kato, T., Haruki, M., Imanaka, T., Morikawa, M., and Kanaya, S. (2001). Isolation and characterization of psychrotrophic bacteria from oil-reservoir and oil sands. <i>Applied Microbial Biotechnology.</i> 55, 794-800. |
- | Add 3.3µL of 6x loading dye to each reaction mixture and load 10µL final volume onto a 1% agarose (in TAE) gel.<br>
| + | <br> |
- | Add 1µL of 6x loading dye to 1µL of GeneRuler 1kb ladder (at 0.5µg/µL)<br>
| + | Prozorov, T., Mallapragada, S. K., Narasimhan, B., Wang, L., Palo, P., Nilsen-Hamilton, M., Williams, T. J., Bazylinski, D. A., Prozorov, R., and Canfield, P. C. (2007). Protein-mediated synthesis of uniform superparamagnetic magnetite nanocrystals. Adv. Funct. Mater. Advanced Functional Materials. 17, 951-957 |
- | Gel loading order as follows:<br>
| + | |
- | <table><table border="3">
| + | =<font color="white">Sponsors= |
- | <tr><td><b>Lane</b></td><td><b>Sample</b></td></tr>
| + | |
- | <tr><td>1</td><td>1kb Ladder</td></tr> | + | |
- | <tr><td>2</td><td>Tango Control</td></tr> | + | |
- | <tr><td>3</td><td>DpnI (Tango)</td></tr> | + | |
- | <tr><td>4</td><td>BcuI/SpeI (Tango)</td></tr> | + | |
- | <tr><td>5</td><td>XbaI (Tango)</td></tr> | + | |
- | <tr><td>6</td><td>EcoRI (Red)</td></tr>
| + | |
- | <tr><td>7</td><td>PstI (Red)</td></tr> | + | |
- | <tr><td>8</td><td>Red Control</td></tr>
| + | |
- | <tr><td>9</td><td>Empty</td></tr>
| + | |
- | <tr><td>10</td><td>Empty</td></tr> | + | |
- | </table><br>
| + | |
- | Ran gel at 100V for 1 hour<br>
| + | |
- | <b>Results:</b> pUC19 plasmid DNA not present at a high enough concentration to visualize by ethidium bromide staining (1kb ladder did stain). <br>
| + | |
- | <b>Conclusion:</b> Will have to re-run experiment with DNA that is present at high enough concentrations to visualize by ethidium bromide staining<br><br><br>
| + | |
| | | |
| + | ==<font color="white">Platinum== |
| | | |
- | <a name="may"></a>
| + | ===<font color="white">Alberta Innovates Technology Futures=== |
- | <b><font size=+2>May 5/2010</font>(in the lab: JV)</b><br>
| + | <html> |
- | <b>Objective:</b> Test Restriction Endonucleases for activity (take 2)<br>
| + | <center> |
- | <b>Relevant Information:</b><br>
| + | <table> |
- | Plasmid DNA used here will be "ES-pSB-CEYFP" from last year's plasmid stocks<br>
| + | <tr> |
- | Prefix Enzymes are: EcoRI and XbaI<br>
| + | <th> |
- | Suffix Enyzmes are: SpeI and PstI<br>
| + | <a href="http://www.albertainnovates.ca/technology/introduction" target="_blank"> |
- | (JV worked out in lab notebook which buffers would be best for each prefix/suffix enzyme combination)<br>
| + | <img src="https://static.igem.org/mediawiki/2010/d/db/UofLAITFlogo.jpg" width="300"/> |
- | Reactions will be assembled as follows:<br>
| + | </a> |
- | <table><table border="3"> | + | </th> |
- | <tr><td><b>Enzyme</td><td>Buffer</td><td>Volume MM(µL)</td><td>Volume Enzyme(µL)</td></tr></b> | + | </tr> |
- | <tr><td>PstI</td><td>Red</td><td>19.75</td><td>.25</td></tr> | + | |
- | <tr><td>XbaI</td><td>Tango</td><td>19.75</td><td>.25</td></tr> | + | |
- | <tr><td>SpeI</td><td>Tango</td><td>19.75</td><td>.25</td></tr>
| + | |
- | <tr><td>EcoRI</td><td>Red</td><td>19.75</td><td>.25</td></tr>
| + | |
- | <tr><td>EcoRI/SpeI</td><td>Red</td><td>19.5</td><td>.25+.25</td></tr>
| + | |
- | <tr><td>XbaI/SpeI</td><td>Tango</td><td>19.5</td><td>.25+.25</td></tr>
| + | |
- | <tr><td>EcoRI/PstI</td><td>Red</td><td>19.5</td><td>.25+.25</td></tr>
| + | |
- | <tr><td>XbaI/PstI</td><td>Tango</td><td>19.5</td><td>.25+.25</td></tr>
| + | |
- | </table><br>
| + | |
- | Make up Master Mixes as follows:<br>
| + | |
- | <table><table border="3">
| + | |
- | <tr><td><b>Red MM</td><td>per tube(µL)</td><td>Total*(µL)</td></tr></b> | + | |
- | <tr><td>MilliQ H<sub>2</sub>0</td><td>15.75</td><td>86.675</td></tr>
| + | |
- | <tr><td>Red Buffer (10x)</td><td>2</td><td>11</td></tr>
| + | |
- | <tr><td>pDNA**</td><td>2</td><td>11</td></tr>
| + | |
- | </table><br>
| + | |
- | <table><table border="3">
| + | |
- | <tr><td><b>Tango MM</td><td>per tube(µL)</td><td>Total*(µL)</td></tr></b>
| + | |
- | <tr><td>MilliQ H<sub>2</sub>0</td><td>15.75</td><td>86.675</td></tr>
| + | |
- | <tr><td>Tango Buffer (10x)</td><td>2</td><td>11</td></tr>
| + | |
- | <tr><td>pDNA**</td><td>2</td><td>11</td></tr>
| + | |
| </table> | | </table> |
- | *Volume per reaction multiplied by 5.5<br>
| + | </center> |
- | **Unknown concentration of pDNA<br><br>
| + | </html> |
- | Incubated for 70min at 37<sup>o</sup>C (Start-1:05pm; End-2:15pm)<br>
| + | |
- | Added 3.3µL of 6x loading dye to each reaction mixture and loaded 10µL onto a 1% agarose gel (in TAE)<br>
| + | ===<font color="white">Oil Sands Initiative=== |
- | Added 1µL of 6x loading dye to 2µL of gene ruler 1kb ladder<br>
| + | <b>https://2010.igem.org/Oil_Sands</b> |
- | Load order as follows:<br>
| + | |
- | <table><table border="3">
| + | |
- | <tr><td><b>Lane</td><td>Sample</td><td>Volume Loaded (µL)</td></tr></b>
| + | |
- | <tr><td>1</td><td>pSB-CEYFP/PstI</td><td>10</td></tr>
| + | |
- | <tr><td>2</td><td>pSB-CEYFP/EcoRI</td><td>10</td></tr>
| + | |
- | <tr><td>3</td><td>pSB-CEYFP/EcoRI/PstI</td><td>10</td></tr>
| + | |
- | <tr><td>4</td><td>pSB-CEYFP/EcoRI/SpeI</td><td>10</td></tr>
| + | |
- | <tr><td>5</td><td>pSB-CEYFP/XbaI/PstI</td><td>10</td></tr>
| + | |
- | <tr><td>6</td><td>pSB-CEYFP/XbaI</td><td>10</td></tr>
| + | |
- | <tr><td>7</td><td>pSB-CEYFP/SpeI</td><td>10</td></tr>
| + | |
- | <tr><td>8</td><td>pSB-CEYFP/XbaI/SpeI</td><td>10</td></tr>
| + | |
- | <tr><td>9</td><td>pSB-CEYFP/Red Master Mix Control</td><td>10</td></tr>
| + | |
- | <tr><td>10</td><td>pSB-CEYFP/Tango Master Mix Control</td><td>10</td></tr>
| + | |
- | <tr><td>11</td><td>pSB-CEYFP/MilliQ H<sub>2</sub>0 Control</td><td>10</td></tr>
| + | |
- | <tr><td>12</td><td>Ladder</td><td>4</td></tr>
| + | |
- | </table><br>
| + | |
- | Ran gel at 100V for 1 hour<br><br>
| + | |
- | <b>Results:</b><br>
| + | |
- | <a href="https://static.igem.org/mediawiki/2010/c/c3/100505JV-EnzymeTest1Cropped.jpg"><img src="https://static.igem.org/mediawiki/2010/c/c3/100505JV-EnzymeTest1Cropped.jpg" width="225" height="151"/></a><br>
| + | |
- | This gel shows that SpeI does not cut on its own, and does not cut when combined with other enzymes<br>
| + | |
- | <b>Conclusion:</b> Test other source of SpeI to see if it has any activity.<br><br>
| + | |
| | | |
- | <b><font size=+2>May 6/2010</font>(in the lab:KG, AS)</b><br> | + | <html> |
- | <b>Objective:</b> To check if the old SpeI enzyme (exp date: March 2011) will cleave plasmid DNA, since we believe the newer SpeI enzyme (exp date: 2012) does not.<br> | + | <center> |
- | <b>Method:</b><br>
| + | <table border="0" cellpadding="5" cellspacing="5" width="40%"> |
- | <table><table border ="3">
| + | <tr> |
- | <tr><td><b>Red Master Mix</b></td><td>per tube (µL)</td><td>Total Volume*</td></tr> | + | <th> |
- | <tr><td>MilliQ H<sub>2</sub>0 Water</td><td>15.75</td><td>63</td></tr> | + | <a href="http://www.see.ualberta.ca/OSRIN.cfm" target="_blank"> |
- | <tr><td>Red Buffer (10x)</td><td>2</td><td>8</td></tr>
| + | <img src="https://static.igem.org/mediawiki/2010/a/a1/UofLOSRINlogo.jpg" width="250"/> |
- | <tr><td>pDNA**</td><td>2</td><td>8</td></tr> | + | </a> |
| + | </th> |
| + | <th><a href="http://www.iseee.ca" target="_blank"> |
| + | <img src="https://static.igem.org/mediawiki/2010/6/6c/UofLISEEElogo.jpg" width="550"/> |
| + | </a></th> |
| </table> | | </table> |
- | *Volume per tube multiplied by 4<br>
| + | </center> |
- | **Used pSB NEYFP pDNA from cell E5 in plasmid box<br>
| + | </html> |
- | Enzymes that will use Red Master Mix are: EcoRI+SpeI (old), EcoRI+SpeI (new)<br>
| + | |
- | Add 0.25µL of each enzyme to 19.5µL of master mix<br><br>
| + | <html> |
- | <table><table border ="3">
| + | <center> |
- | <tr><td><b>Tango Master Mix</b></td><td>per tube (µL)</td><td>Total Volume*</td></tr> | + | <table border="0" cellpadding="5" cellspacing="5" width="95%"> |
- | <tr><td>MilliQ H<sub>2</sub>0 Water</td><td>15.75</td><td>94.5</td></tr> | + | <tr> |
- | <tr><td>Tango Buffer (10x)</td><td>2</td><td>12</td></tr> | + | <th> |
- | <tr><td>pDNA**</td><td>2</td><td>12</td></tr> | + | <a href="http://www.conocophillips.ca" target="_blank"> |
| + | <img src="https://static.igem.org/mediawiki/2010/4/42/UofLConocoPhillipslogo.jpg" width="160"/> |
| + | </a> |
| + | </th> |
| + | <th><a href="http://www.nexeninc.com" target="_blank"> |
| + | <img src="https://static.igem.org/mediawiki/2010/2/2e/UofLNexenlogo.jpg" width="120"/> |
| + | </a> |
| + | </th> |
| + | <th><a href="http://www.statoil.com" target="_blank"> |
| + | <img src="https://static.igem.org/mediawiki/2010/4/45/UofLStatoillogo.jpg" width="90"/> |
| + | </a> |
| + | </th> |
| + | <th><a href="http://www.suncor.com" target="_blank"> |
| + | <img src="https://static.igem.org/mediawiki/2010/c/c3/UofLSuncorlogo.jpg" width="130"/> |
| + | </a> |
| + | </th> |
| + | <th><a href="http://www.total-ep-canada.com" target="_blank"> |
| + | <img src="https://static.igem.org/mediawiki/2010/e/ed/UofLTotallogo.jpg" width="75"/> |
| + | </a> |
| + | </th> |
| </table> | | </table> |
- | *Volume per tube multiplied by 6<br>
| + | </center> |
- | **Used pSB NEYFP pDNA from cell E5 in plasmid box<br>
| + | </html> |
- | Enzymes that will use Tango Master Mix are: SpeI (old), SpeI (new), XbaI+SpeI (old), XbaI+SpeI (new)<br>
| + | |
- | Add 0.25µL of each enzyme to 19.5µL of master mix<br><br>
| + | |
- | Incubated all reactions at 37<sup>o</sup>C for 1h (Start-8:30pm; End-9:30pm)<br>
| + | |
- | Will not be able to run on agarose gel tonight, will label them so JV can run them in the morning<br>
| + | |
- | <u>Tube Names:</u><br>
| + | |
- | Master Mix 1 Control (Red Buffer)<br>
| + | |
- | Master Mix 2 Control (Tango Buffer)<br>
| + | |
- | E+S(N); EcoRI + SpeI(N)<br>
| + | |
- | E+S(O); EcoRI + SpeI(O)<br>
| + | |
- | X+S(N); XbaI + SpeI(N)<br>
| + | |
- | X+S(O); XbaI + SpeI(O)<br>
| + | |
- | S(N); SpeI(N)<br>
| + | |
- | S(O); SpeI(O)<br><br>
| + | |
- | Placed in -20<sup>o</sup>C freezer of later analysis by agarose electrophoresis<br><br>
| + | |
| | | |
| + | ==<font color="white">Gold== |
| | | |
- | <b><font size=+2>May 10/2010</font>(in the lab:JV)</b><br>
| + | |
- | <b>Objective:</b> To analyze the restriction test done by KG and AS on May 6/2010 by agarose electrophoresis<br><br> | + | ===<font color="white">Autodesk=== |
- | <b>Method:</b><br> | + | <html> |
- | <table><table border="3"> | + | <center> |
- | <tr><td><b>Lane</b></td><td>Sample</td><td>Quantity Loaded (µL)</td></tr> | + | <table> |
- | <tr><td>1</td><td>MM1 Control</td><td>10</td></tr> | + | <tr> |
- | <tr><td>2</td><td>MM2 Control</td><td>10</td></tr> | + | <th> |
- | <tr><td>3</td><td>EcoRI+SpeI(N)</td><td>10</td></tr> | + | <a href="http://www.autodeskresearch.com/" target="_blank"> |
- | <tr><td>4</td><td>EcoRI+SpeI(O)</td><td>10</td></tr>
| + | <img src="https://static.igem.org/mediawiki/2010/6/67/UofLAutodesklogo.jpg"/> |
- | <tr><td>5</td><td>SpeI(N)</td><td>10</td></tr>
| + | </a> |
- | <tr><td>6</td><td>SpeI(O)</td><td>10</td></tr>
| + | </th> |
- | <tr><td>7</td><td>XbaI+SpeI(N)</td><td>10</td></tr>
| + | </tr> |
- | <tr><td>8</td><td>XbaI+SpeI(O)</td><td>10</td></tr>
| + | |
- | <tr><td>9</td><td>1kb Ladder</td><td>5</td></tr>
| + | |
| </table> | | </table> |
- | Run gel for 60min at 100V<br><br>
| + | </center> |
- | <b>Results:</b><br>
| + | </html> |
- | <a href="https://static.igem.org/mediawiki/2010/b/b0/100510JRV-EnzymeTest1Cropped.jpg"><img src="https://static.igem.org/mediawiki/2010/b/b0/100510JRV-EnzymeTest1Cropped.jpg" width="225" height="151"/></a><br> | + | |
- | It appears as though both SpeI enzymes are working properly here. We will utilize the newer batch of SpeI (expires 2012) from this point forward.<br><br>
| + | |
| | | |
| | | |
- | <b><font size=+2>May 10/2010</font>(in the lab:JV, KG, AV)</b><br>
| + | ==<font color="white">Silver== |
- | <b>Objective:</b>Make 24 LB agar plates with 100µg/mL ampicillin antibiotic.<br>
| + | |
- | <b>Method:</b>Make 2L of LB media with agar<br>
| + | |
- | 2x10g Tryptone<br>
| + | |
- | 2X2.5g Yeast Extract<br>
| + | |
- | 2x5g NaCl<br>
| + | |
- | 2x10g Agar<br><br>
| + | |
- | Continued <b> May 11/2010<br></b>
| + | |
- | (Stock Ampicillin solution is 100mg/mL)<br>
| + | |
- | Have 4x500mL of LB with Agar<br>
| + | |
- | Add 500µL of stock ampicillin to 500mL of media<br><br>
| + | |
- | <b><font size=+2>May 11/2010 Evening </font> (in the lab: KG, AV, MC, TF, JV, JS)<br></b>
| + | |
- | <b>Objective:</b> To transform the following plasmids into DH5α <i>E.coli</i> cells.
| + | |
- | <table><table border="3">
| + | |
- | <tr><td><b>Construct Name (2009)</td></b><td><b>Construct Location (2009)</b></td></tr>
| + | |
- | <tr><td>Lumazine</td><td>J4</td></tr>
| + | |
- | <tr><td>Lumazine-dT</td><td>J5,J6</td></tr>
| + | |
- | <tr><td>sRBS-Lumazine-dT</td><td>J7,J8</td></tr>
| + | |
- | <tr><td>pBAD-TetR</td><td>I4</td></tr>
| + | |
- | <tr><td>pBAD</td><td>A5,F10</td></tr>
| + | |
- | <tr><td>sRBS</td><td>D5,E10</td></tr>
| + | |
- | <tr><td>pSB-CEYFP</td><td>E5,D6</td></tr>
| + | |
- | <tr><td>pSB-NEYFP</td><td>F5,C6</td></tr>
| + | |
- | <tr><td>C-term Tag</td><td>C10</td></tr>
| + | |
- | <tr><td>N-term Tag</td><td>D9,D10</td></tr>
| + | |
- | <tr><td>pTet</td><td>E4</td></tr>
| + | |
- | <tr><td>EYFP</td><td>A4</td></tr>
| + | |
- | <tr><td>CFP Complete</td><td>D4</td></tr>
| + | |
- | </table><br>
| + | |
- | <b>Method: </b>Followed <a href="https://2010.igem.org/Team:Lethbridge#transformation">"Competent Cell Transformation"</a> protocol in Common Protocols section and plated on LB agar supplemented with ampicillin.<br>
| + | |
- | <b>Results: </b>The following plasmids were successfully transformed and formed colonies:<br>
| + | |
- | <ul>
| + | |
- | <li>Lumazine (J4)</li>
| + | |
- | <li>sRBS-Lumazine-dT (J7)</li>
| + | |
- | <li>sRBS-Lumazine-dT (J8)</li>
| + | |
- | <li>pBAD (A5) </li>
| + | |
- | <li>pBAD (F10) </li>
| + | |
- | <li>pSB-CEYFP </li>
| + | |
- | <li>pSB-NEYFP </li>
| + | |
- | <li>N-term tag </li>
| + | |
- | <li>EYFP (A4)</li>
| + | |
- | <li>CFP Complete (D4)</li></ul><br>
| + | |
- | <b>Conclusion:</b> Need another attempt to transform the following plasmids:<br>
| + | |
- | <ul>
| + | |
- | <li>Lumazine-dT (J5,J6)</li>
| + | |
- | <li>pBAD-TetR </li>
| + | |
- | <li>sRBS (D5,E10)</li>
| + | |
- | <li>C-Term tag</li>
| + | |
- | <li>pTet</li></ul><br>
| + | |
| | | |
- | <br>May 13<br>
| |
| | | |
- | </div> | + | ===<font color="white">MathWorks=== |
- | <div class="TabbedPanelsContent">
| + | <html> |
- | <b><font size=+2>Common Protocols:</b></font><br> | + | <center> |
- | <ol><li><a href=#transformation>Competent Cell Transformation</a></li><br>
| + | <table> |
- | </ol>
| + | <tr> |
- | <a name="transformation"></a>
| + | <th> |
- | <b>Competent Cell Transformation</b><br> | + | <a href="http://www.mathworks.com/" target="_blank"> |
- | <ol>
| + | <img src="https://static.igem.org/mediawiki/2010/c/c3/UofLMathWorkslogo.jpg" width="300" border="0"/> |
- | <li>Thaw 20µL of aliquotted cells (DH5α of BL21(DE3)) on ice.</li>
| + | </a> |
- | <li>Gently pipet 2.0µL of DNA into competent cells<br>
| + | </th> |
- | ATTENTION: <br>
| + | </tr> |
- | Do not perform any additional mixing<br>
| + | </table> |
- | Never use more DNA that <u>10% of the volume of the competent cells </u>otherwise the cells get destroyed by osmotic shock</li>
| + | </center> |
- | <li>Incubate the cells on ice for 30 minutes.</li>
| + | </html> |
- | <li>Heat shock the cells in a water bath at <u>42<sup>o</sup>C for EXACTLY 45 seconds.</u></li>
| + | |
- | <li>Incubate the cells on ice for 1 minute.</li>
| + | |
- | <li>Add 250µL sterile media to the cells and incubate at 37<sup>o</sup>C for 1 hour with shaking (200RPM).</li>
| + | |
- | <li>Plate 100µL and 50µL on prewarmed LB agar plate containing the appropriate antibiotic.<br>
| + | |
- | For ligations, plate all 250µL.</li>
| + | |
- | <li>Leave plate for 10-15 minutes to soak the cell suspension into the agar.</li>
| + | |
- | <li>Flip plate over (agar on top)</li>
| + | |
- | <li>Incubate the plates in the 37<sup>o</sup>C incubator overnight</li>
| + | |
- | </ol> | + | |
| | | |
- | </div>
| |
- | </div>
| |
- | </div>
| |
| | | |
| + | ===<font color="white">Geneious=== |
| + | <html> |
| + | <center> |
| + | <table> |
| + | <tr> |
| + | <th> |
| + | <a href="http://www.geneious.com/" target="_blank"> |
| + | <img src="https://static.igem.org/mediawiki/2010/3/30/UofLGeneiouslogo.jpg" width="300"/> |
| + | </a> |
| + | </th> |
| + | </tr> |
| + | </table> |
| + | </center> |
| + | </html> |
| | | |
- | </div>
| + | |
- | <div class="TabbedPanelsContent"><iframe src="https://www.google.com/calendar/embed?showPrint=0&height=800&wkst=1&bgcolor=%2333ff33&src=uleth.igem%40gmail.com&color=%2329527A&ctz=America%2FEdmonton" style=" border-width:0 " width="962" height="800" frameborder="0" scrolling="no"></iframe></div>
| + | ==<font color="white">Bronze== |
- | <div class="TabbedPanelsContent">Content 6</div>
| + | |
- | <div class="TabbedPanelsContent">Content 7</div>
| + | ===<font color="white">Integrated DNA Technologies=== |
- | </div>
| + | <html> |
- | </div> | + | <center> |
- | <p> </p> | + | <table> |
- | <p> </p>
| + | <tr> |
- | <p>This site is best viewed with fire fox</p>
| + | <th> |
- | <script type="text/javascript"> | + | <a href="http://www.idtdna.com/" target="_blank"> |
- | <!--
| + | <img src="https://static.igem.org/mediawiki/2010/4/43/UofLIDTlogo.jpg"/> |
- | var TabbedPanels1 = new Spry.Widget.TabbedPanels("TabbedPanels1");
| + | </a> |
- | var TabbedPanels2 = new Spry.Widget.TabbedPanels("TabbedPanels2");
| + | </th> |
- | var TabbedPanels3 = new Spry.Widget.TabbedPanels("TabbedPanels3");
| + | </tr> |
- | var TabbedPanels4 = new Spry.Widget.TabbedPanels("TabbedPanels4");
| + | </table> |
- | //--> | + | </center> |
- | </script> | + | |
- | </body> | + | |
| </html> | | </html> |
| + | |
| + | |
| + | ===<font color="white">GeneArt & Mr. Gene=== |
| + | <html> |
| + | <center> |
| + | <align="left"> |
| + | <table border="0" cellpadding="5" cellspacing="5" width="40%"> |
| + | <tr> |
| + | <th> |
| + | <a href="http://www.geneart.com/" target="_blank"> |
| + | <img src="https://static.igem.org/mediawiki/2010/e/ed/UofLGENEARTlogo.jpg"/> |
| + | </a> |
| + | </th> |
| + | <th><a href="http://mrgene.com/desktopdefault.aspx/tabid-2/" target="_blank"> |
| + | <img src="https://static.igem.org/mediawiki/2010/2/26/UofLMRGENElogo.jpg"/> |
| + | </a></th> |
| + | </table> |
| + | </center> |
| + | </html> |
| + | |
| + | |
| + | ===<font color="white">Macrogen=== |
| + | <html> |
| + | <center> |
| + | <table> |
| + | <tr> |
| + | <th> |
| + | <a href="http://www.macrogen.com/eng/macrogen/macrogen_main.jsp" target="_blank"> |
| + | <img src="https://static.igem.org/mediawiki/2010/a/a7/UofLMacrogenlogo.jpg"/> |
| + | </a> |
| + | </th> |
| + | </tr> |
| + | </table> |
| + | </center> |
| + | </html> |
| + | |
| + | ==<font color="white">Sponsorship Breakdown== |
| + | ===<font color="white">The University of Lethbridge iGEM Team requests your support in the following ways:=== |
| + | |
| + | a) $2000 scholarship of one team member – It is our hope that every team member will be able to travel to the iGEM competition taking place in November 2010. However, without enough funding, this will not be the situation. By sponsoring one student at a time, we can work towards reaching our goal of the entire U of L team traveling to MIT and provide every student with the possibility to meet and talk to some of the brightest and inspiring minds of synthetic biology. |
| + | |
| + | b) Monetary donation towards lab supplies, travel arrangements and iGEM competition registration fees. |
| + | |
| + | c) Tangible donations of lab supplies or other necessary materials. |
| + | |
| + | ===<font color="white">Donations can be made in two ways:=== |
| + | |
| + | 1. Donations can be directed towards the U of L iGEM Scholarship account. |
| + | |
| + | 2. Donations can be directed towards the U of L iGEM General account and in recognition of your generous support, the U of L iGEM team would like to recognize you and/or your business as outlined in the sponsorship levels. |
| + | |
| + | ===<font color="white">Sponsorship Levels=== |
| + | |
| + | Platinum - $5000+ or gift in kind <br> |
| + | Logo on team shirts, large logo on scientific poster, large logo on team wiki and verbal recognition during team project presentations/media interviews. |
| + | |
| + | Gold - $2000-$4999 or gift in kind<br> |
| + | Logo on team shirts, medium logo on scientific poster, medium logo on team wiki and written recognition at end of team project presentations. |
| + | |
| + | Silver - $1000-$1999 or gift in kind<br> |
| + | Medium logo on team shirts, scientific poster, medium logo on team wiki and written recognition at end of team project presentations. |
| + | |
| + | Bronze - <$999 or gift in kind<br> |
| + | Small logo on team shirts, scientific poster, small logo on team wiki and written recognition at end of team project presentations. |
| + | <br> |
| + | <br> |
| + | <br> |