http://2012.igem.org/wiki/index.php?title=Team:Macquarie_Australia/Notebook&feed=atom&action=historyTeam:Macquarie Australia/Notebook - Revision history2024-03-29T09:03:07ZRevision history for this page on the wikiMediaWiki 1.16.0http://2012.igem.org/wiki/index.php?title=Team:Macquarie_Australia/Notebook&diff=283144&oldid=prevRyankenny at 10:47, 26 October 20122012-10-26T10:47:58Z<p></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><div class="accordionButton"><div id="protocol">POST-REGIONAL JAMBOREE: <del class="diffchange diffchange-inline">Wednesday 26th September</del></div></div></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><div class="accordionButton"><div id="protocol">POST-REGIONAL JAMBOREE: <ins class="diffchange diffchange-inline">Tuesday 23rd October</ins></div></div></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <img src="https://static.igem.org/mediawiki/2012/7/77/KAIST_Protocol_content_top.png" id="top-part"/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <img src="https://static.igem.org/mediawiki/2012/7/77/KAIST_Protocol_content_top.png" id="top-part"/></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <div id="protocolcontent"></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <div id="protocolcontent"></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>Today, <del class="diffchange diffchange-inline">Rob and Harry used </del>the <del class="diffchange diffchange-inline">French press </del>to <del class="diffchange diffchange-inline">successfully lyse </del>the <del class="diffchange diffchange-inline">BL21 cells that had previously been transformed</del>. <del class="diffchange diffchange-inline">These </del>were <del class="diffchange diffchange-inline">then spun down </del>and the <del class="diffchange diffchange-inline">supernatant was analysed using the Nanodrop</del>. <del class="diffchange diffchange-inline">The 1C-3C constructs displayed absorbance peaks at 400nm </del>and <del class="diffchange diffchange-inline">700nm, indicating </del>the <del class="diffchange diffchange-inline">coupling of </del>the bacteriophytochrome <del class="diffchange diffchange-inline">with the chromophore. The absorbance peak at 700nm was very wide, indicating that a percentage of the bacteriophytochrome construct had been photoconverted </del>to the far-red <del class="diffchange diffchange-inline">conformation</del>. The <del class="diffchange diffchange-inline">4K constructs that did not contain biliverdin only gave </del>the <del class="diffchange diffchange-inline">expected absorbance peak for protein solution at 280nm</del>. <del class="diffchange diffchange-inline"></p></del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>Today, <ins class="diffchange diffchange-inline">Ryan performed </ins>the <ins class="diffchange diffchange-inline">last of the experiments </ins>to <ins class="diffchange diffchange-inline">get </ins>the <ins class="diffchange diffchange-inline">switch reversing</ins>. <ins class="diffchange diffchange-inline">Cultures </ins>were <ins class="diffchange diffchange-inline">grown up containing the Agro BioBrick </ins>and the <ins class="diffchange diffchange-inline">composite part we produced</ins>. <ins class="diffchange diffchange-inline">Cells were harvested </ins>and <ins class="diffchange diffchange-inline">washed in </ins>the <ins class="diffchange diffchange-inline">dark to prevent </ins>the bacteriophytochrome <ins class="diffchange diffchange-inline">from converting </ins>to the far-red <ins class="diffchange diffchange-inline">form</ins>. The <ins class="diffchange diffchange-inline">cells were lysed using a French press to produce </ins>the <ins class="diffchange diffchange-inline">cell lysate, and the free bacteriophytochrome</ins>. <ins class="diffchange diffchange-inline">A UV-Vis spectra was obtained for the uninduced </ins>and <ins class="diffchange diffchange-inline">induced culture </ins>with <ins class="diffchange diffchange-inline">and without biliverdin added. A significantly more detailed </ins>analysis of the <ins class="diffchange diffchange-inline">spectra obtained can be seen in </ins>the <ins class="diffchange diffchange-inline">Results section </ins><a href="https://2012.igem.org/Team:Macquarie_Australia/<ins class="diffchange diffchange-inline">Results#switch</ins>">here</a>. <ins class="diffchange diffchange-inline">The spectra obtained can be seen below,</p></ins></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"><p>Rob </del>and <del class="diffchange diffchange-inline">Harry continued </del>with <del class="diffchange diffchange-inline">the spectral </del>analysis of the <del class="diffchange diffchange-inline">BL21 cell samples yielded </del>the <del class="diffchange diffchange-inline">spectra shown </del><a href="https://2012.igem.org/Team:Macquarie_Australia/<del class="diffchange diffchange-inline">Protocols/spectralanalysis</del>">here</a>.</div></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><center><img src="https://static.igem.org/mediawiki/2012/3/37/4Kspectra.png" width=600 height=360></center></ins></div></div></td></tr>
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</table>Ryankennyhttp://2012.igem.org/wiki/index.php?title=Team:Macquarie_Australia/Notebook&diff=282998&oldid=prevRyankenny at 10:19, 26 October 20122012-10-26T10:19:49Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Today, Rob and Harry used the French press to successfully lyse the BL21 cells that had previously been transformed. These were then spun down and the supernatant was analysed using the Nanodrop. The 1C-3C constructs displayed absorbance peaks at 400nm and 700nm, indicating the coupling of the bacteriophytochrome with the chromophore. The absorbance peak at 700nm was very wide, indicating that a percentage of the bacteriophytochrome construct had been photoconverted to the far-red conformation. The 4K constructs that did not contain biliverdin only gave the expected absorbance peak for protein solution at 280nm. </p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Today, Rob and Harry used the French press to successfully lyse the BL21 cells that had previously been transformed. These were then spun down and the supernatant was analysed using the Nanodrop. The 1C-3C constructs displayed absorbance peaks at 400nm and 700nm, indicating the coupling of the bacteriophytochrome with the chromophore. The absorbance peak at 700nm was very wide, indicating that a percentage of the bacteriophytochrome construct had been photoconverted to the far-red conformation. The 4K constructs that did not contain biliverdin only gave the expected absorbance peak for protein solution at 280nm. </p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Rob and Harry continued with the spectral analysis of the BL21 cell samples yielded the spectra shown <a href="https://2012.igem.org/Team:Macquarie_Australia/Protocols/spectralanalysis">here</a>.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Rob and Harry continued with the spectral analysis of the BL21 cell samples yielded the spectra shown <a href="https://2012.igem.org/Team:Macquarie_Australia/Protocols/spectralanalysis">here</a>.</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><div class="accordionButton"><div id="protocol">POST-REGIONAL JAMBOREE: Wednesday 26th September</div></div></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><div class="accordionButton"><div id="protocol">POST-REGIONAL JAMBOREE: Wednesday 26th September</div></div></div></td></tr>
</table>Ryankennyhttp://2012.igem.org/wiki/index.php?title=Team:Macquarie_Australia/Notebook&diff=282985&oldid=prevRyankenny at 10:18, 26 October 20122012-10-26T10:18:36Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>To browse through our notebook simply click on the image with the date. It will expand with what we did that week. To continue on simply click on the next one. If the notebook isn't functioning as planned (it will not load the next tab of content) then please go to our linearised view which is located <a href="https://2012.igem.org/Team:Macquarie_Australia/Notebook2">here</a>.</p><br><br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>To browse through our notebook simply click on the image with the date. It will expand with what we did that week. To continue on simply click on the next one. If the notebook isn't functioning as planned (it will not load the next tab of content) then please go to our linearised view which is located <a href="https://2012.igem.org/Team:Macquarie_Australia/Notebook2">here</a>.</p><br><br></div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;"> </del></div></td><td colspan="2"> </td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <div class="accordionButton" id="open"><div id="protocol">Week 1- Tuesday July 31st</div></div></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <div class="accordionButton" id="open"><div id="protocol">Week 1- Tuesday July 31st</div></div></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><div class="accordionButton"><div id="protocol">Week 2- Tuesday 7th August</div></div></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><div class="accordionButton"><div id="protocol">Week 2- Tuesday 7th August</div></div></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Today, Rob and Harry used the French press to successfully lyse the BL21 cells that had previously been transformed. These were then spun down and the supernatant was analysed using the Nanodrop. The 1C-3C constructs displayed absorbance peaks at 400nm and 700nm, indicating the coupling of the bacteriophytochrome with the chromophore. The absorbance peak at 700nm was very wide, indicating that a percentage of the bacteriophytochrome construct had been photoconverted to the far-red conformation. The 4K constructs that did not contain biliverdin only gave the expected absorbance peak for protein solution at 280nm. </p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Today, Rob and Harry used the French press to successfully lyse the BL21 cells that had previously been transformed. These were then spun down and the supernatant was analysed using the Nanodrop. The 1C-3C constructs displayed absorbance peaks at 400nm and 700nm, indicating the coupling of the bacteriophytochrome with the chromophore. The absorbance peak at 700nm was very wide, indicating that a percentage of the bacteriophytochrome construct had been photoconverted to the far-red conformation. The 4K constructs that did not contain biliverdin only gave the expected absorbance peak for protein solution at 280nm. </p></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>Rob and Harry continued with the spectral analysis of the BL21 cell samples yielded the spectra shown <a href="https://2012.igem.org/Team:Macquarie_Australia/Protocols/spectralanalysis">here</a>.<del class="diffchange diffchange-inline"></div></del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>Rob and Harry continued with the spectral analysis of the BL21 cell samples yielded the spectra shown <a href="https://2012.igem.org/Team:Macquarie_Australia/Protocols/spectralanalysis">here</a>.</div></td></tr>
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</table>Ryankennyhttp://2012.igem.org/wiki/index.php?title=Team:Macquarie_Australia/Notebook&diff=282979&oldid=prevRyankenny at 10:16, 26 October 20122012-10-26T10:16:20Z<p></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> </del><div class="accordionButton"><div id="protocol"><del class="diffchange diffchange-inline">Week 9</del>- Wednesday 26th September</div></div></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><div class="accordionButton"><div id="protocol"><ins class="diffchange diffchange-inline">POST</ins>-<ins class="diffchange diffchange-inline">REGIONAL JAMBOREE: </ins>Wednesday 26th September</div></div></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <img src="https://static.igem.org/mediawiki/2012/7/77/KAIST_Protocol_content_top.png" id="top-part"/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <img src="https://static.igem.org/mediawiki/2012/7/77/KAIST_Protocol_content_top.png" id="top-part"/></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <div id="protocolcontent"></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <div id="protocolcontent"></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><p>Today, Rob and Harry used the French press to successfully lyse the BL21 cells that had previously been transformed. These were then spun down and the supernatant was analysed using the Nanodrop. The 1C-3C constructs displayed absorbance peaks at 400nm and 700nm, indicating the coupling of the bacteriophytochrome with the chromophore. The absorbance peak at 700nm was very wide, indicating that a percentage of the bacteriophytochrome construct had been photoconverted to the far-red conformation. The 4K constructs that did not contain biliverdin only gave the expected absorbance peak for protein solution at 280nm. </p></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><p>Rob and Harry continued with the spectral analysis of the BL21 cell samples yielded the spectra shown <a href="https://2012.igem.org/Team:Macquarie_Australia/Protocols/spectralanalysis">here</a>.</div></ins></div></td></tr>
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</table>Ryankennyhttp://2012.igem.org/wiki/index.php?title=Team:Macquarie_Australia/Notebook&diff=282967&oldid=prevRyankenny at 10:13, 26 October 20122012-10-26T10:13:47Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>The SDS PAGE Gel under IR light indicates the successful coupling of biliverdin with bacteriophytochromes. Biliverdin acts as a fluorophore, accepting a photon of a particular wavelength and emitting at a longer wavelength. We can see three bands in the second gel which indicates that an infrared active molecule has bound to heme oxygenase. For a more indepth analysis of the gel see <a href="https://2012.igem.org/Team:Macquarie_Australia/Results#gel">here</a>.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>The SDS PAGE Gel under IR light indicates the successful coupling of biliverdin with bacteriophytochromes. Biliverdin acts as a fluorophore, accepting a photon of a particular wavelength and emitting at a longer wavelength. We can see three bands in the second gel which indicates that an infrared active molecule has bound to heme oxygenase. For a more indepth analysis of the gel see <a href="https://2012.igem.org/Team:Macquarie_Australia/Results#gel">here</a>.</p></div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> </del><div class="accordionButton"><div id="protocol">Week 9- Wednesday 26th September</div></div></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> </ins></div></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <img src="https://static.igem.org/mediawiki/2012/7/77/KAIST_Protocol_content_top.png" id="top-part"/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <img src="https://static.igem.org/mediawiki/2012/7/77/KAIST_Protocol_content_top.png" id="top-part"/></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Today, Rob and Harry used the French press to successfully lyse the BL21 cells that had previously been transformed. These were then spun down and the supernatant was analysed using the Nanodrop. The 1C-3C constructs displayed absorbance peaks at 400nm and 700nm, indicating the coupling of the bacteriophytochrome with the chromophore. The absorbance peak at 700nm was very wide, indicating that a percentage of the bacteriophytochrome construct had been photoconverted to the far-red conformation. The 4K constructs that did not contain biliverdin only gave the expected absorbance peak for protein solution at 280nm. </p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Today, Rob and Harry used the French press to successfully lyse the BL21 cells that had previously been transformed. These were then spun down and the supernatant was analysed using the Nanodrop. The 1C-3C constructs displayed absorbance peaks at 400nm and 700nm, indicating the coupling of the bacteriophytochrome with the chromophore. The absorbance peak at 700nm was very wide, indicating that a percentage of the bacteriophytochrome construct had been photoconverted to the far-red conformation. The 4K constructs that did not contain biliverdin only gave the expected absorbance peak for protein solution at 280nm. </p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Rob and Harry continued with the spectral analysis of the BL21 cell samples yielded the spectra shown <a href="https://2012.igem.org/Team:Macquarie_Australia/Protocols/spectralanalysis">here</a>.</div></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Rob and Harry continued with the spectral analysis of the BL21 cell samples yielded the spectra shown <a href="https://2012.igem.org/Team:Macquarie_Australia/Protocols/spectralanalysis">here</a>.</div></div></td></tr>
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<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;"> <div class="accordionButton"><div id="protocol">Week 9- Wednesday 26th September</div></div></ins></div></td></tr>
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</table>Ryankennyhttp://2012.igem.org/wiki/index.php?title=Team:Macquarie_Australia/Notebook&diff=238225&oldid=prevRyankenny at 03:53, 27 September 20122012-09-27T03:53:54Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Today, Rob and Harry used the French press to successfully lyse the BL21 cells that had previously been transformed. These were then spun down and the supernatant was analysed using the Nanodrop. The 1C-3C constructs displayed absorbance peaks at 400nm and 700nm, indicating the coupling of the bacteriophytochrome with the chromophore. The absorbance peak at 700nm was very wide, indicating that a percentage of the bacteriophytochrome construct had been photoconverted to the far-red conformation. The 4K constructs that did not contain biliverdin only gave the expected absorbance peak for protein solution at 280nm. </p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Today, Rob and Harry used the French press to successfully lyse the BL21 cells that had previously been transformed. These were then spun down and the supernatant was analysed using the Nanodrop. The 1C-3C constructs displayed absorbance peaks at 400nm and 700nm, indicating the coupling of the bacteriophytochrome with the chromophore. The absorbance peak at 700nm was very wide, indicating that a percentage of the bacteriophytochrome construct had been photoconverted to the far-red conformation. The 4K constructs that did not contain biliverdin only gave the expected absorbance peak for protein solution at 280nm. </p></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>Rob and Harry continued with the spectral analysis of the BL21 cell samples yielded the spectra shown here.</div></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>Rob and Harry continued with the spectral analysis of the BL21 cell samples yielded the spectra shown <ins class="diffchange diffchange-inline"><a href="https://2012.igem.org/Team:Macquarie_Australia/Protocols/spectralanalysis"></ins>here<ins class="diffchange diffchange-inline"></a></ins>.</div></div></td></tr>
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</table>Ryankennyhttp://2012.igem.org/wiki/index.php?title=Team:Macquarie_Australia/Notebook&diff=238129&oldid=prevRyankenny at 03:52, 27 September 20122012-09-27T03:52:48Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p> Please see our <a href="https://2012.igem.org/Team:Macquarie_Australia/Results">Results</a> page for more information about our gels.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p> Please see our <a href="https://2012.igem.org/Team:Macquarie_Australia/Results">Results</a> page for more information about our gels.</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center><h3>Transformations</h3></center></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center><h3>Transformations</h3></center></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>Using the <a href="https://2012.igem.org/Team:Macquarie/Protocols/TransformationProtocol">standard <del class="diffchange diffchange-inline">trasformation </del>protocol</a> Matt and Ryan transformed BL21 to express T7 Heme Oxygenase, T7 Agro Bacteriophytochrome and the T7 Heme Oxygenase-Deinococcus ligation product. These were cultured onto Kanamycin selective plates and left to incubate at room temperature over the weekend. They would be used to test the function of the two components.</p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>Using the <a href="https://2012.igem.org/Team:Macquarie/Protocols/TransformationProtocol">standard <ins class="diffchange diffchange-inline">transformation </ins>protocol</a> Matt and Ryan transformed BL21 to express T7 Heme Oxygenase, T7 Agro Bacteriophytochrome and the T7 Heme Oxygenase-Deinococcus ligation product. These were cultured onto Kanamycin selective plates and left to incubate at room temperature over the weekend. They would be used to test the function of the two components.</p></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Today, Rob and Harry used the French press to successfully lyse the BL21 cells that had previously been transformed. These were then spun down and the supernatant was analysed using the Nanodrop. The 1C-3C constructs displayed absorbance peaks at 400nm and 700nm, indicating the coupling of the bacteriophytochrome with the chromophore. The absorbance peak at 700nm was very wide, indicating that a percentage of the bacteriophytochrome construct had been photoconverted to the far-red conformation. The 4K constructs that did not contain biliverdin only gave the expected absorbance peak for protein solution at 280nm. </p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Today, Rob and Harry used the French press to successfully lyse the BL21 cells that had previously been transformed. These were then spun down and the supernatant was analysed using the Nanodrop. The 1C-3C constructs displayed absorbance peaks at 400nm and 700nm, indicating the coupling of the bacteriophytochrome with the chromophore. The absorbance peak at 700nm was very wide, indicating that a percentage of the bacteriophytochrome construct had been photoconverted to the far-red conformation. The 4K constructs that did not contain biliverdin only gave the expected absorbance peak for protein solution at 280nm. </p></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div></div></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><p>Rob and Harry continued with the spectral analysis of the BL21 cell samples yielded the spectra shown here.</ins></div></div></td></tr>
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</table>Ryankennyhttp://2012.igem.org/wiki/index.php?title=Team:Macquarie_Australia/Notebook&diff=226784&oldid=prevElleworgan at 00:24, 27 September 20122012-09-27T00:24:14Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><h3><center>Bacteriophytochrome Characterisation</h3></center></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><h3><center>Bacteriophytochrome Characterisation</h3></center></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Ridding on the excellent news that our oxygenase was functional we lysed the cells and ran an SDS PAGE gel to determine if the bacteriophytochrome was being expressed in the case of the BioBrick and if it was coupling with biliverdin for our composite.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Ridding on the excellent news that our oxygenase was functional we lysed the cells and ran an SDS PAGE gel to determine if the bacteriophytochrome was being expressed in the case of the BioBrick and if it was coupling with biliverdin for our composite.</p></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><center><img src="<del class="diffchange diffchange-inline">https</del>://<del class="diffchange diffchange-inline">static</del>.<del class="diffchange diffchange-inline">igem</del>.<del class="diffchange diffchange-inline">org/mediawiki/2012/0/0e</del>/<del class="diffchange diffchange-inline">GEL5MOD</del>.<del class="diffchange diffchange-inline">png</del>" height=400 width=476></center></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><center><img src="<ins class="diffchange diffchange-inline">http</ins>://<ins class="diffchange diffchange-inline">24</ins>.<ins class="diffchange diffchange-inline">media</ins>.<ins class="diffchange diffchange-inline">tumblr.com</ins>/<ins class="diffchange diffchange-inline">tumblr_mazfnjrDIz1rg4kjpo1_500</ins>.<ins class="diffchange diffchange-inline">jpg</ins>" height=400 width=476></center></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>The gel contains (left to right): Agro + T7 (4K) Sample 3, 4K Sample 1, Heme Oxygenase T7 (1C) + Deinococcus (3C) (1C+3C) sample 7, 1C+3C Sample 6, 1C Sample 6, 1C sample 5, 1C3C Sample 3, Marker.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>The gel contains (left to right): Agro + T7 (4K) Sample 3, 4K Sample 1, Heme Oxygenase T7 (1C) + Deinococcus (3C) (1C+3C) sample 7, 1C+3C Sample 6, 1C Sample 6, 1C sample 5, 1C3C Sample 3, Marker.</p></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><center><img src="<del class="diffchange diffchange-inline">https</del>://<del class="diffchange diffchange-inline">static</del>.<del class="diffchange diffchange-inline">igem</del>.<del class="diffchange diffchange-inline">org/mediawiki/2012/2/22</del>/<del class="diffchange diffchange-inline">GEL6MOD</del>.<del class="diffchange diffchange-inline">png</del>" height=400 width=476></center></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><center><img src="<ins class="diffchange diffchange-inline">http</ins>://<ins class="diffchange diffchange-inline">24</ins>.<ins class="diffchange diffchange-inline">media</ins>.<ins class="diffchange diffchange-inline">tumblr.com</ins>/<ins class="diffchange diffchange-inline">tumblr_mazfliiW7O1rg4kjpo1_500</ins>.<ins class="diffchange diffchange-inline">jpg</ins>" height=400 width=476></center></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>This gel contains (left to right): 1C3C Sample 3 Induced, 1C3C Sample 5 Induced, 4K1 Induced, 4K3 Induced, 1C3C Sample 3 Uninduced, 1C3C Sample 5 Uninduced, 1C3C Sample 7, 4K1 Uninduced, 4K3 Uninduced, Marker.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>This gel contains (left to right): 1C3C Sample 3 Induced, 1C3C Sample 5 Induced, 4K1 Induced, 4K3 Induced, 1C3C Sample 3 Uninduced, 1C3C Sample 5 Uninduced, 1C3C Sample 7, 4K1 Uninduced, 4K3 Uninduced, Marker.</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><h3><center>The Switch</center></h3></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><h3><center>The Switch</center></h3></div></td></tr>
</table>Elleworganhttp://2012.igem.org/wiki/index.php?title=Team:Macquarie_Australia/Notebook&diff=225758&oldid=prevRyankenny at 00:04, 27 September 20122012-09-27T00:04:49Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><tr><td>Agrobacterium</td></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><tr><td>Agrobacterium</td></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><td>Xba1 and Pst1</td></tr></table><br></center></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><td>Xba1 and Pst1</td></tr></table><br></center></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"><p> Please see our <a href="https://2012.igem.org/Team:Macquarie_Australia/Results">Results</a> page for more information about our gels.</p></del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center><h3>Characterisation</h3></center></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center><h3>Characterisation</h3></center></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>The Biobricks we had produced were characterised by running on a gel following the restriction digest. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>The Biobricks we had produced were characterised by running on a gel following the restriction digest. </div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>The lanes with expected bands are shown in the 1st and 3rd lanes of this Agro T7 treatment. </li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>The lanes with expected bands are shown in the 1st and 3rd lanes of this Agro T7 treatment. </li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>Linearized backbones </li></ol></blockquote></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>Linearized backbones </li></ol></blockquote></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><p> Please see our <a href="https://2012.igem.org/Team:Macquarie_Australia/Results">Results</a> page for more information about our gels.</p></ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center><h3>Heme Oxygenase Functionality</h3></center></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center><h3>Heme Oxygenase Functionality</h3></center></div></td></tr>
</table>Ryankennyhttp://2012.igem.org/wiki/index.php?title=Team:Macquarie_Australia/Notebook&diff=225610&oldid=prevRyankenny at 00:02, 27 September 20122012-09-27T00:02:15Z<p></p>
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<td colspan='2' style="background-color: white; color:black;">Revision as of 00:02, 27 September 2012</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><tr><td>Agrobacterium</td></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><tr><td>Agrobacterium</td></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><td>Xba1 and Pst1</td></tr></table><br></center></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><td>Xba1 and Pst1</td></tr></table><br></center></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><p> Please see our <a href="https://2012.igem.org/Team:Macquarie_Australia/Results">Results</a> page for more information about our gels.</p></ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center><h3>Characterisation</h3></center></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center><h3>Characterisation</h3></center></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>The Biobricks we had produced were characterised by running on a gel following the restriction digest. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>The Biobricks we had produced were characterised by running on a gel following the restriction digest. </div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Gel 2: From left to right, this gel contains digests of: Mw ladder, T7-Heme Oxygenase (1C), Agro Bacteriophytochrome(5C), T7 Agrobacteriophytochrome (4C), Heme Oxygenase (2C), Kanamycin vector, chloramphenicol vector, 1C+5C ligation product in kanamycin vector, 4K+2K in chloramphenicol.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Gel 2: From left to right, this gel contains digests of: Mw ladder, T7-Heme Oxygenase (1C), Agro Bacteriophytochrome(5C), T7 Agrobacteriophytochrome (4C), Heme Oxygenase (2C), Kanamycin vector, chloramphenicol vector, 1C+5C ligation product in kanamycin vector, 4K+2K in chloramphenicol.</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>The gels provided substantial evidence that the ligation was successful and that we had produced the composite product.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>The gels provided substantial evidence that the ligation was successful and that we had produced the composite product.</p></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;"><p> Please see our <a href="https://2012.igem.org/Team:Macquarie_Australia/Results">Results</a> page for more information about our gels.</p></ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center><h3>Transformations</h3></center></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center><h3>Transformations</h3></center></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Using the <a href="https://2012.igem.org/Team:Macquarie/Protocols/TransformationProtocol">standard trasformation protocol</a> Matt and Ryan transformed BL21 to express T7 Heme Oxygenase, T7 Agro Bacteriophytochrome and the T7 Heme Oxygenase-Deinococcus ligation product. These were cultured onto Kanamycin selective plates and left to incubate at room temperature over the weekend. They would be used to test the function of the two components.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Using the <a href="https://2012.igem.org/Team:Macquarie/Protocols/TransformationProtocol">standard trasformation protocol</a> Matt and Ryan transformed BL21 to express T7 Heme Oxygenase, T7 Agro Bacteriophytochrome and the T7 Heme Oxygenase-Deinococcus ligation product. These were cultured onto Kanamycin selective plates and left to incubate at room temperature over the weekend. They would be used to test the function of the two components.</p></div></td></tr>
</table>Ryankenny