http://2007.igem.org/wiki/index.php?title=Melb:Plan:GFP_Fluorescent_reporter&feed=atom&action=history
Melb:Plan:GFP Fluorescent reporter - Revision history
2024-03-29T10:58:29Z
Revision history for this page on the wiki
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http://2007.igem.org/wiki/index.php?title=Melb:Plan:GFP_Fluorescent_reporter&diff=14532&oldid=prev
Patricia at 07:55, 11 August 2007
2007-08-11T07:55:02Z
<p></p>
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<td colspan='2' style="background-color: white; color:black;">Revision as of 07:55, 11 August 2007</td>
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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;">[[Melbourne/Plan|<back to plans>]]</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 style="color: red; font-weight: bold; text-decoration: none;"></ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==LacZ reporter==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==LacZ reporter==</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Ligate LacZ coding region to gentamycin RBS part. (Done?)(1 day)</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Ligate LacZ coding region to gentamycin RBS part. (Done?)(1 day)</div></td></tr>
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Patricia
http://2007.igem.org/wiki/index.php?title=Melb:Plan:GFP_Fluorescent_reporter&diff=14531&oldid=prev
Patricia: /* LacZ reporter */
2007-08-11T07:53:52Z
<p><span class="autocomment">LacZ reporter</span></p>
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<td colspan='2' style="background-color: white; color:black;">Revision as of 07:53, 11 August 2007</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Liquid culture, miniprep and screen via restriction digest (EcoRI and SpeI), digest double terminator (P1 1G) simultaneously (EcoRI and XbaI) (overnight + 1 day)</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Liquid culture, miniprep and screen via restriction digest (EcoRI and SpeI), digest double terminator (P1 1G) simultaneously (EcoRI and XbaI) (overnight + 1 day)</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Make glycerol stock of RBS-LacZ containing cells. (same day)</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Make glycerol stock of RBS-LacZ containing cells. (same day)</div></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;"></del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Ligate the RBS-LacZ construct into double terminator plasmid. (same day)</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Ligate the RBS-LacZ construct into double terminator plasmid. (same day)</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Transform ligation and select for construct using Gen/Amp plate. (1 day + overnight)</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Transform ligation and select for construct using Gen/Amp plate. (1 day + overnight)</div></td></tr>
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Patricia
http://2007.igem.org/wiki/index.php?title=Melb:Plan:GFP_Fluorescent_reporter&diff=14530&oldid=prev
Patricia at 07:53, 11 August 2007
2007-08-11T07:53:37Z
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<td colspan='2' style="background-color: white; color:black;">Revision as of 07:53, 11 August 2007</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>This construct can then be ligated to appropriate promoters for use as a test harness</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>This construct can then be ligated to appropriate promoters for use as a test harness <ins class="diffchange diffchange-inline">with the substrate 3,4-cyclohexenoesculetin--D-galactopyranoside (S-Gal). For information on conditions see supplementary info from the following paper:</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">Nature 438, 441-442 (24 November 2005)</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">Synthetic biology: Engineering Escherichia coli to see light</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">Anselm Levskaya1, Aaron A. Chevalier2, Jeffrey J. Tabor2, Zachary Booth Simpson2, Laura A. Lavery2, Matthew Levy2, Eric A. Davidson2, Alexander Scouras2, Andrew D. Ellington2,3, Edward M. Marcotte2,3 and Christopher A. Voigt1,4,5</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> </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">Test harnesses needed:</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">*OmpR promoter: Ligate to promoter (3 days) and express in Envz+ cells under osmotic stress (time?)</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">*Red light system: Above construct in Envz- cells with red light system</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">*Blue light system: Ligate to Promoter responding to ComA (3 days), in Envz- cells constitutively expressing ComA and Blue light receptors (time?)</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">*AND gate: Use above construct in Envz- cells constitutively expressing Blue light receptor and Red light receptor with ComA expression under the control of OmpR promoter. (time?)</ins></div></td></tr>
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Patricia
http://2007.igem.org/wiki/index.php?title=Melb:Plan:GFP_Fluorescent_reporter&diff=14529&oldid=prev
Patricia: /* LacZ reporter */
2007-08-11T07:41:19Z
<p><span class="autocomment">LacZ reporter</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 07:41, 11 August 2007</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==LacZ reporter==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==LacZ reporter==</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#Ligate LacZ coding region to gentamycin RBS part. (Done?)</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Ligate LacZ coding region to gentamycin RBS part. (Done?<ins class="diffchange diffchange-inline">)(1 day</ins>)</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#Transform and select for appropriate construct using Kan/Gen plate. (Done?)</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Transform and select for appropriate construct using Kan/Gen plate. (Done?<ins class="diffchange diffchange-inline">)(1 day + overnight</ins>)</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#Liquid culture and screen via restriction digest (EcoRI and SpeI), digest double terminator (P1 1G) simultaneously <del class="diffchange diffchange-inline"> </del>(EcoRI and XbaI) <del class="diffchange diffchange-inline"> </del>(<del class="diffchange diffchange-inline">Monday 13/08/07</del>)</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Liquid culture<ins class="diffchange diffchange-inline">, miniprep </ins>and screen via restriction digest (EcoRI and SpeI), digest double terminator (P1 1G) simultaneously (EcoRI and XbaI) <ins class="diffchange diffchange-inline"> </ins>(<ins class="diffchange diffchange-inline">overnight + 1 day</ins>)</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#Make glycerol stock of RBS-LacZ containing cells. (<del class="diffchange diffchange-inline">Monday 13/08/07</del>)</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Make glycerol stock of RBS-LacZ containing cells. (<ins class="diffchange diffchange-inline">same day</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>#Ligate the RBS-LacZ construct into double terminator plasmid. (<del class="diffchange diffchange-inline">Monday 13/08/07</del>)</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Ligate the RBS-LacZ construct into double terminator plasmid. (<ins class="diffchange diffchange-inline">same day</ins>)</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#Transform ligation and select for construct </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Transform ligation and select for construct <ins class="diffchange diffchange-inline">using Gen/Amp plate. (1 day + overnight)</ins></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">Liquid culture, miniprep and confirm using EcoRI/Pst digest (overnight + 1 day)</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">#prepare glycerol stock (same day)</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> </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">This construct can then be ligated to appropriate promoters for use as a test harness</ins></div></td></tr>
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Patricia
http://2007.igem.org/wiki/index.php?title=Melb:Plan:GFP_Fluorescent_reporter&diff=14528&oldid=prev
Patricia at 07:29, 11 August 2007
2007-08-11T07:29:17Z
<p></p>
<p><b>New page</b></p><div>==LacZ reporter==<br />
#Ligate LacZ coding region to gentamycin RBS part. (Done?)<br />
#Transform and select for appropriate construct using Kan/Gen plate. (Done?)<br />
#Liquid culture and screen via restriction digest (EcoRI and SpeI), digest double terminator (P1 1G) simultaneously (EcoRI and XbaI) (Monday 13/08/07)<br />
#Make glycerol stock of RBS-LacZ containing cells. (Monday 13/08/07)<br />
<br />
#Ligate the RBS-LacZ construct into double terminator plasmid. (Monday 13/08/07)<br />
#Transform ligation and select for construct <br />
#</div>
Patricia