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Refactored Decaffeination Operon - Revision history
2024-03-29T09:16:14Z
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http://2012.igem.org/wiki/index.php?title=Refactored_Decaffeination_Operon&diff=75978&oldid=prev
Erik.quandt: /* Experiment 2: Construction of a refactored decaffeination operon */
2012-08-20T19:42:04Z
<p><span class="autocomment">Experiment 2: Construction of a refactored decaffeination operon</span></p>
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<td colspan='2' style="background-color: white; color:black;">← Older revision</td>
<td colspan='2' style="background-color: white; color:black;">Revision as of 19:42, 20 August 2012</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Run 5 uL on .8% agarose gel<br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Run 5 uL on .8% agarose gel<br/></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>[[File:2L.jpg|200px]]</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>[[File:2L.jpg|200px]]<ins class="diffchange diffchange-inline"><br/></ins></div></td></tr>
<tr><td colspan="2"> </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><ins class="diffchange diffchange-inline">Lanes: <br/></ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>1. 2k1<br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>1. 2k1<br/></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>2. 2k2<br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>2. 2k2<br/></div></td></tr>
</table>
Erik.quandt
http://2012.igem.org/wiki/index.php?title=Refactored_Decaffeination_Operon&diff=75977&oldid=prev
Erik.quandt at 19:41, 20 August 2012
2012-08-20T19:41:12Z
<p></p>
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<td colspan='2' style="background-color: white; color:black;">← Older revision</td>
<td colspan='2' style="background-color: white; color:black;">Revision as of 19:41, 20 August 2012</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Run 5 uL on .8% agarose gel<br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Run 5 uL on .8% agarose gel<br/></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">insert gel image (on darkroom computer?)</del><br/></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">[[File:2L.jpg|200px]]</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">1. 2k1</ins><br/></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">2. 2k2<br/></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">3. GeneRuler 1kb ladder</ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Result: both clones show anticipated ~5kB band corresponding to the complete operon.<br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Result: both clones show anticipated ~5kB band corresponding to the complete operon.<br/></div></td></tr>
</table>
Erik.quandt
http://2012.igem.org/wiki/index.php?title=Refactored_Decaffeination_Operon&diff=75956&oldid=prev
Erik.quandt: /* Experiment 2: Construction of a refactored decaffeination operon */
2012-08-20T19:23:51Z
<p><span class="autocomment">Experiment 2: Construction of a refactored decaffeination operon</span></p>
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<td colspan='2' style="background-color: white; color:black;">← Older revision</td>
<td colspan='2' style="background-color: white; color:black;">Revision as of 19:23, 20 August 2012</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>dilute 60 uL 2g transformations (1-2) -> 3mL Mineral M9 media + 34ug/mL chloramphenicol +/- 100mg Caffeine or Theophylline. Place on 30 deg shaker x 48 hrs<br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>dilute 60 uL 2g transformations (1-2) -> 3mL Mineral M9 media + 34ug/mL chloramphenicol +/- 100mg Caffeine or Theophylline. Place on 30 deg shaker x 48 hrs<br/></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Results (growth):<del class="diffchange diffchange-inline"><br/></del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Results (growth):</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">{| cellpadding="5" border="1" cellspacing="0"</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">! Media</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">! 2g1 (+operon)</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">! 2g2 (vector only)</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">|-</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">| M9</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">| -</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">| -</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">|-</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">| M9 + Caffeine</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">| -</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">| -</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">|-</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">| M9 + Theophylline</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">| +</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">| -</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">|-</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">|}</ins></div></td></tr>
<tr><td colspan="2"> </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;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<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;"> 2g1 (+operon) 2g2 (vector only)</del></div></td><td colspan="2"> </td></tr>
<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;">1. M9 - -</del></div></td><td colspan="2"> </td></tr>
<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;">2. M9 + caffeine - -</del></div></td><td colspan="2"> </td></tr>
<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;">3. M9 + Theophylline + _</del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Result: construct enables growth (demethylation) on Theophylline. NdmC is not fuctional.<br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Result: construct enables growth (demethylation) on Theophylline. NdmC is not fuctional.<br/></div></td></tr>
</table>
Erik.quandt
http://2012.igem.org/wiki/index.php?title=Refactored_Decaffeination_Operon&diff=75940&oldid=prev
Erik.quandt: /* Experiment 2: Constructon of a refactored decaffeination operon */
2012-08-20T18:37:47Z
<p><span class="autocomment">Experiment 2: Constructon of a refactored decaffeination operon</span></p>
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<td colspan='2' style="background-color: white; color:black;">Revision as of 18:37, 20 August 2012</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>==Experiment 2: <del class="diffchange diffchange-inline">Constructon </del>of a refactored decaffeination operon==</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>==Experiment 2: <ins class="diffchange diffchange-inline">Construction </ins>of a refactored decaffeination operon==</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>2a. Phusion PCR<br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>2a. Phusion PCR<br/></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Run 1 uL 2a1-7, 3.5 uL Fermentas 1kb <del class="diffchange diffchange-inline">Generuler </del>ladder</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Run 1 uL 2a1-7, 3.5 uL Fermentas 1kb <ins class="diffchange diffchange-inline">GeneRuler </ins>ladder</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[File:2a.jpg]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[File:2a.jpg]]</div></td></tr>
</table>
Erik.quandt
http://2012.igem.org/wiki/index.php?title=Refactored_Decaffeination_Operon&diff=75939&oldid=prev
Erik.quandt: /* Experiment 2: Constructon of a refactored decaffeination operon */
2012-08-20T18:35:51Z
<p><span class="autocomment">Experiment 2: Constructon of a refactored decaffeination operon</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Experiment 2: Constructon of a refactored decaffeination operon==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Experiment 2: Constructon of a refactored decaffeination operon==</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
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Erik.quandt
http://2012.igem.org/wiki/index.php?title=Refactored_Decaffeination_Operon&diff=75938&oldid=prev
Erik.quandt: /* Experiment 2: Constructon of a refactored decaffeination operon */
2012-08-20T18:34:58Z
<p><span class="autocomment">Experiment 2: Constructon of a refactored decaffeination operon</span></p>
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<td colspan='2' style="background-color: white; color:black;">Revision as of 18:34, 20 August 2012</td>
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<td colspan="2" class="diff-lineno">Line 54:</td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">Gel igm2a<br/></del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">Run 1 uL 2a1-7, 3.5 uL Fermentas 1kb Generuler ladder</ins></div></td></tr>
<tr><td colspan="2"> </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><ins class="diffchange diffchange-inline">[[File:2a.jpg]]</ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td colspan="2" class="diff-lineno">Line 156:</td>
<td colspan="2" class="diff-lineno">Line 158:</td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>50 deg thermocycler x 60 minutes<br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>50 deg thermocycler x 60 minutes<br/></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Run 5 uL each reaction on .8% agarose gel<br/></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Run 5 uL each reaction <ins class="diffchange diffchange-inline">(1-4)</ins>on .8% agarose gel <ins class="diffchange diffchange-inline">+ 3.5 ul Fermentas 1kb GeneRuler</ins><br/></div></td></tr>
<tr><td colspan="2"> </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><ins class="diffchange diffchange-inline">[[File:2f.jpg]]</ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<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;">insert igm2f.jpeg<br/></del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Desalted gibson reactions on .025uM Nitrocellulose membranes x 20 minutes<br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Desalted gibson reactions on .025uM Nitrocellulose membranes x 20 minutes<br/></div></td></tr>
</table>
Erik.quandt
http://2012.igem.org/wiki/index.php?title=Refactored_Decaffeination_Operon&diff=75925&oldid=prev
Erik.quandt at 18:28, 20 August 2012
2012-08-20T18:28:18Z
<p></p>
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Erik.quandt
http://2012.igem.org/wiki/index.php?title=Refactored_Decaffeination_Operon&diff=75879&oldid=prev
Erik.quandt at 17:52, 20 August 2012
2012-08-20T17:52:59Z
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Erik.quandt
http://2012.igem.org/wiki/index.php?title=Refactored_Decaffeination_Operon&diff=75875&oldid=prev
Erik.quandt at 17:51, 20 August 2012
2012-08-20T17:51:30Z
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Erik.quandt
http://2012.igem.org/wiki/index.php?title=Refactored_Decaffeination_Operon&diff=75869&oldid=prev
Erik.quandt at 17:49, 20 August 2012
2012-08-20T17:49:43Z
<p></p>
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Erik.quandt