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Team:Groningen/Notebook/24 August 2009 - Revision history
2024-03-28T13:28:53Z
Revision history for this page on the wiki
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http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/24_August_2009&diff=53661&oldid=prev
Nienke: /* Metal Accumulation */
2009-08-25T16:48:49Z
<p><span class="autocomment">Metal Accumulation</span></p>
<table style="background-color: white; color:black;">
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<td colspan='2' style="background-color: white; color:black;">Revision as of 16:48, 25 August 2009</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: #eee; color:black; font-size: smaller;"><div>*'''Cloning of MymT into pSB1AC3 or other vectors has stopped'''</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*'''Cloning of MymT into pSB1AC3 or other vectors has stopped'''</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;">**It is considered to be to much time consuming to also clone MymT next to the other MTs.</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>===Vectors===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Vectors===</div></td></tr>
</table>
Nienke
http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/24_August_2009&diff=53658&oldid=prev
Nienke: /* Metal Accumulation */
2009-08-25T16:47:20Z
<p><span class="autocomment">Metal Accumulation</span></p>
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<td colspan='2' style="background-color: white; color:black;">Revision as of 16:47, 25 August 2009</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>**Thereby the construct of Mymt-pSB1AC3 nr7-9 do not give correct fragment size, also the constructs pSB1AC3-SmtA nr 1 and pSB1AC3-fMT nr 5 do not give correct fragment size.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>**Thereby the construct of Mymt-pSB1AC3 nr7-9 do not give correct fragment size, also the constructs pSB1AC3-SmtA nr 1 and pSB1AC3-fMT nr 5 do not give correct fragment size.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>**Put o/n culture @ warm room of pSB1AC3-SmtA nr2&3 and pSB1AC3-fMT nr 4&6.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>**Put o/n culture @ warm room of pSB1AC3-SmtA nr2&3 and pSB1AC3-fMT nr 4&6.</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 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;">*'''Cloning of MymT into pSB1AC3 or other vectors has stopped'''</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>===Vectors===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Vectors===</div></td></tr>
</table>
Nienke
http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/24_August_2009&diff=53653&oldid=prev
Nienke: /* Metal Accumulation */
2009-08-25T16:33:23Z
<p><span class="autocomment">Metal Accumulation</span></p>
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<td colspan='2' style="background-color: white; color:black;">Revision as of 16:33, 25 August 2009</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Image:F102471 2009-08-24 Colony PCR fMT SmtA MymT.JPG|350px]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Image:F102471 2009-08-24 Colony PCR fMT SmtA MymT.JPG|350px]]</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>**From left to right: </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>**From left to right: </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>::Upper: SmtA-1 till 3, fMT 4-6, MymT 7-8, AC3 (using VF2/VR primers)</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>::Upper: SmtA-1 till 3 <ins class="diffchange diffchange-inline">(1200bp)</ins>, fMT 4-6 <ins class="diffchange diffchange-inline">(500bp)</ins>, MymT 7-8<ins class="diffchange diffchange-inline">(500)</ins>, AC3 <ins class="diffchange diffchange-inline">(315bp) </ins>(using VF2/VR primers)</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>::Lower: SmtA-1 till 3, fMT 4-6, MymT 7-9 (using gene specific primers)</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>::Lower: SmtA-1 till 3 <ins class="diffchange diffchange-inline">(900bp)</ins>, fMT 4-6 <ins class="diffchange diffchange-inline">(200bp)</ins>, MymT 7-9<ins class="diffchange diffchange-inline">(215bp) </ins>(using gene specific primers)</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">**Thereby the construct of Mymt-pSB1AC3 nr7-9 do not give correct fragment size, also the constructs pSB1AC3-SmtA nr 1 and pSB1AC3-fMT nr 5 do not give correct fragment size.</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">**Put o/n culture @ warm room of pSB1AC3-SmtA nr2&3 and pSB1AC3-fMT nr 4&6.</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>===Vectors===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Vectors===</div></td></tr>
</table>
Nienke
http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/24_August_2009&diff=53615&oldid=prev
Nienke: /* Metal Accumulation */
2009-08-25T15:09:35Z
<p><span class="autocomment">Metal Accumulation</span></p>
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<td colspan='2' style="background-color: white; color:black;">Revision as of 15:09, 25 August 2009</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: #eee; color:black; font-size: smaller;"><div>===Metal Accumulation===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Metal Accumulation===</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;">*'''Check pSB1AC3-mymT, fmt, smtA'''</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;">**Pick 3 colonies of ''E. coli'' TOP10 + pSB1AC3-mymT, fmt, smtA.</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;">**Add 1uL MQ and microwave 1min to lyse the cells.</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;">**Do colony PCR with VR/VF primers and RBS-fw/gene specific rev primers.</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;">**Run PCR products on gel.</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;">[[Image:F102471 2009-08-24 Colony PCR fMT SmtA MymT.JPG|350px]]</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;">**From left to right: </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;">::Upper: SmtA-1 till 3, fMT 4-6, MymT 7-8, AC3 (using VF2/VR primers)</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;">::Lower: SmtA-1 till 3, fMT 4-6, MymT 7-9 (using gene specific primers)</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>===Vectors===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Vectors===</div></td></tr>
</table>
Nienke
http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/24_August_2009&diff=52891&oldid=prev
Wilfred: /* Metal promotors */
2009-08-24T15:40:37Z
<p><span class="autocomment">Metal promotors</span></p>
<table style="background-color: white; color:black;">
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<td colspan='2' style="background-color: white; color:black;">Revision as of 15:40, 24 August 2009</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>** Plasmids that utilize the same replication system cannot co-exist in the same bacterial cell.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>** Plasmids that utilize the same replication system cannot co-exist in the same bacterial cell.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>** pMB1<sup>*</sup> and ColE1 are in different [http://wolfson.huji.ac.il/expression/vector/origin.html#comp compatibility group]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>** pMB1<sup>*</sup> and ColE1 are in different [http://wolfson.huji.ac.il/expression/vector/origin.html#comp compatibility group]</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;">** pMB1 and ColE1 are too similar and thus not compatible</ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><sup>*</sup> pMB1 is the pUC19 derived ori present in pSB1AC3</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><sup>*</sup> pMB1 is the pUC19 derived ori present in pSB1AC3</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>
</table>
Wilfred
http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/24_August_2009&diff=52882&oldid=prev
Verhoeven1981: /* GVP Cluster */
2009-08-24T15:37:42Z
<p><span class="autocomment">GVP Cluster</span></p>
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<td colspan='2' style="background-color: white; color:black;">Revision as of 15:37, 24 August 2009</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Image:24-8 no.1.jpg|400px]] [[Image:Generulers_1kb_marker_Fermentas.jpg]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Image:24-8 no.1.jpg|400px]] [[Image:Generulers_1kb_marker_Fermentas.jpg]]</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>:→ From left to right: 1kB ladder, <del class="diffchange diffchange-inline">Ars</del>, <del class="diffchange diffchange-inline">Cue</del>, <del class="diffchange diffchange-inline">Znt</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>:→ From left to right: 1kB ladder, <ins class="diffchange diffchange-inline">pArsR (4x)</ins>, <ins class="diffchange diffchange-inline">pCueO (3x)</ins>, <ins class="diffchange diffchange-inline">pZntR (3x)</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">:→ The concentration of plasmids was not determined before restriction was done, and likely the concentration was to high to give a band at ~3000bp of uncut plasmid.</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">[[Image:24-8 no.2.jpg|700px]]</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">:→ From left to right: 1kB ladder, pArsR (2x), pCueO (2x), pZntR (2x), Control BBa_J23101, Empty Slot</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">:→ The 2% Agarose gel was run for 15 min. at 100V to get separation at low bp fragments, larger parts are grouped together at the top</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">:→ The expected sizes are seen in the gel, plasmids can be send for sequencing!</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>===Transporters===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Transporters===</div></td></tr>
</table>
Verhoeven1981
http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/24_August_2009&diff=52858&oldid=prev
Wilfred: /* Metal promotors */
2009-08-24T15:25:35Z
<p><span class="autocomment">Metal promotors</span></p>
<table style="background-color: white; color:black;">
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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 15:25, 24 August 2009</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* Plasmid compatibility: its ability to replicate in conjuction with another plasmid.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* Plasmid compatibility: its ability to replicate in conjuction with another plasmid.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>** Plasmids that utilize the same replication system cannot co-exist in the same bacterial cell.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>** Plasmids that utilize the same replication system cannot co-exist in the same bacterial cell.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>** <del class="diffchange diffchange-inline">pUC19 </del>and ColE1 are in different [http://wolfson.huji.ac.il/expression/vector/origin.html#comp compatibility group]</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>** <ins class="diffchange diffchange-inline">pMB1<sup>*</sup> </ins>and ColE1 are in different [http://wolfson.huji.ac.il/expression/vector/origin.html#comp compatibility group]</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"><sup>*</sup> pMB1 is the pUC19 derived ori present in pSB1AC3</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>==Dry==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Dry==</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>{{Team:Groningen/Notebook/Day/Footer}}</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{Team:Groningen/Notebook/Day/Footer}}</div></td></tr>
</table>
Wilfred
http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/24_August_2009&diff=52823&oldid=prev
Verhoeven1981: /* GVP Cluster */
2009-08-24T14:52:28Z
<p><span class="autocomment">GVP Cluster</span></p>
<table style="background-color: white; color:black;">
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<td colspan='2' style="background-color: white; color:black;">Revision as of 14:52, 24 August 2009</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Image:24-8 no.1.jpg|400px]] [[Image:Generulers_1kb_marker_Fermentas.jpg]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Image:24-8 no.1.jpg|400px]] [[Image:Generulers_1kb_marker_Fermentas.jpg]]</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>:→ From left to right: 1kB ladder,</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>:→ From left to right: 1kB ladder, <ins class="diffchange diffchange-inline">Ars, Cue, Znt</ins></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Transporters===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Transporters===</div></td></tr>
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Verhoeven1981
http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/24_August_2009&diff=52822&oldid=prev
Verhoeven1981: /* GVP Cluster */
2009-08-24T14:51:00Z
<p><span class="autocomment">GVP Cluster</span></p>
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<td colspan='2' style="background-color: white; color:black;">Revision as of 14:51, 24 August 2009</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>:→ {{todo}} Test promoter strenght compared to BBa_J23101 promoter (Sven)</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>:→ {{todo}} Test promoter strenght compared to BBa_J23101 promoter (Sven)</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>:→ {{todo}} Enter sequences of constructs to Sandbox</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>:→ {{todo}} Enter sequences of constructs to Sandbox</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 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 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;">[[Image:Plasmid Miniprep Kit Flow Protocol.gif|thumb|150px| www.sigmaaldrich.com]]</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 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;">'''Plasmid Purification'''</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 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;">Plasmid isolation was performed on the cultures of E.coli TOP10 containing plasmids [http://partsregistry.org/wiki/index.php?title=Part:BBa_J61002 BBa_J61002] with [http://partsregistry.org/wiki/index.php?title=Part:BBa_K190016 Zinc+RBS], [http://partsregistry.org/wiki/index.php?title=Part:BBa_K190017 Copper+RBS] and [http://partsregistry.org/wiki/index.php?title=Part:BBa_K190015 Arsenic+RBS] promoters and RFP with the "Sygma-Aldrich™ [http://www.sigmaaldrich.com/life-science/molecular-biology/dna-and-rna-purification/plasmid-miniprep-kit.html GenElute™] Plasmid Miniprep Kit".</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 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;">* From each tube 4mL of culture was collected in a 2.0mL cup, and the cells were pelleted by centrifugation for 1 min. at max. speed and the supernatant discarded.</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;">* Plasmids were eluted with 30μL MQ and stored in the fridge</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 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;">'''Restriction Control'''</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 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;">Isolated plasmids were cut with fast-digest enzymes EcoRI and PstI to cut out promoter-RFP, and create ~2000bp and ~950bp fragments.</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 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;">[[Image:24-8 no.1.jpg|400px]] [[Image:Generulers_1kb_marker_Fermentas.jpg]]</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 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;">:→ From left to right: 1kB ladder,</ins></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Transporters===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Transporters===</div></td></tr>
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Verhoeven1981
http://2009.igem.org/wiki/index.php?title=Team:Groningen/Notebook/24_August_2009&diff=52749&oldid=prev
Wilfred: /* Metal promotors */
2009-08-24T13:26:16Z
<p><span class="autocomment">Metal promotors</span></p>
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<td colspan='2' style="background-color: white; color:black;">Revision as of 13:26, 24 August 2009</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Vectors===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Vectors===</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Metal promotors====</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Metal promotors====</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">Protocol </del>#<del class="diffchange diffchange-inline">3</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">[http://wolfson.huji.ac.il/expression/vector/origin.html</ins>#<ins class="diffchange diffchange-inline">top origin of replication] determines:</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">Phosphorylation and annealing synthetic oligo's</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>* <ins class="diffchange diffchange-inline">Vector copy number</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">Ligation </del>in <del class="diffchange diffchange-inline">pSB1AC3</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>* <ins class="diffchange diffchange-inline">Plasmid compatibility: its ability to replicate </ins>in <ins class="diffchange diffchange-inline">conjuction with another plasmid.</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">** Plasmids that utilize the same replication system cannot co-exist in the same bacterial cell.</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">** pUC19 and ColE1 are in different [http://wolfson.huji.ac.il/expression/vector/origin.html#comp compatibility group]</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>==Dry==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Dry==</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{Team:Groningen/Notebook/Day/Footer}}</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{Team:Groningen/Notebook/Day/Footer}}</div></td></tr>
</table>
Wilfred