http://2014.igem.org/wiki/index.php?title=Team:Aachen/Notebook/Wetlab/August&feed=atom&action=historyTeam:Aachen/Notebook/Wetlab/August - Revision history2024-03-28T13:21:46ZRevision history for this page on the wikiMediaWiki 1.16.5http://2014.igem.org/wiki/index.php?title=Team:Aachen/Notebook/Wetlab/August&diff=362965&oldid=prevFgohr: /* 6th */2014-10-17T22:39:29Z<p><span class="autocomment">6th</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* transformation of J04450 in pSB1K3 and pSB1A3 in NEB10β cells.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* transformation of J04450 in pSB1K3 and pSB1A3 in NEB10β cells.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>* <del class="diffchange diffchange-inline">They also did a </del>plasmid prep of J04450 in pSB1C3 and <del class="diffchange diffchange-inline">Flo's vectors</del>.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>* plasmid prep of J04450 in pSB1C3<ins class="diffchange diffchange-inline">, pSEVA234_LasR </ins>and <ins class="diffchange diffchange-inline">pSEVA641_BsFbFP</ins>.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* made precultures of NEB10β and DH5α cells</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* made precultures of NEB10β and DH5α cells</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* inoculation of the fermenter at 11:40, and induced the fermentation of pET17-Gal3. The fermentation is expected to run 24&nbsp;h.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* inoculation of the fermenter at 11:40, and induced the fermentation of pET17-Gal3. The fermentation is expected to run 24&nbsp;h.</div></td></tr>
</table>Fgohrhttp://2014.igem.org/wiki/index.php?title=Team:Aachen/Notebook/Wetlab/August&diff=359111&oldid=prevVeraA: /* 5th */2014-10-17T21:59:24Z<p><span class="autocomment">5th</span></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>{{Team:Aachen/Figure|Aachen_14-07-5_PCR_toREACh1%262.png|title=Agarose gel with PCR products after <del class="diffchange diffchange-inline">Quichchange </del>from K1319000 to K1319001 and K1319002 |width=400px}}</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>{{Team:Aachen/Figure|Aachen_14-07-5_PCR_toREACh1%262.png|title=Agarose gel with PCR products after <ins class="diffchange diffchange-inline">Quikchange </ins>from K1319000 to K1319001 and K1319002 |width=400px}}</div></td></tr>
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</table>VeraAhttp://2014.igem.org/wiki/index.php?title=Team:Aachen/Notebook/Wetlab/August&diff=331195&oldid=prevStefanReinhold: /* 6th */2014-10-17T16:17:56Z<p><span class="autocomment">6th</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* transformation of J04450 in pSB1K3 and pSB1A3 in NEB10β cells.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* transformation of J04450 in pSB1K3 and pSB1A3 in NEB10β cells.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* They also did a plasmid prep of J04450 in pSB1C3 and Flo's vectors.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* They also did a plasmid prep of J04450 in pSB1C3 and Flo's vectors.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>* made precultures of <del class="diffchange diffchange-inline">NEB10βand DH5 alpha </del>cells</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>* made precultures of <ins class="diffchange diffchange-inline">NEB10β and DH5α </ins>cells</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* inoculation of the fermenter at 11:40, and induced the fermentation of pET17-Gal3. The fermentation is expected to run 24&nbsp;h.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* inoculation of the fermenter at 11:40, and induced the fermentation of pET17-Gal3. The fermentation is expected to run 24&nbsp;h.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* as the first Quickchange PCR for REACh2 was not sucsessful, it was reapeted as a gradient PCR with annealing temperatire 55°C, 57°C and 60°C. Then the agarose gel with PCR product samples was made.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* as the first Quickchange PCR for REACh2 was not sucsessful, it was reapeted as a gradient PCR with annealing temperatire 55°C, 57°C and 60°C. Then the agarose gel with PCR product samples was made.</div></td></tr>
</table>StefanReinholdhttp://2014.igem.org/wiki/index.php?title=Team:Aachen/Notebook/Wetlab/August&diff=331169&oldid=prevStefanReinhold: /* 4th */2014-10-17T16:17:35Z<p><span class="autocomment">4th</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>== 4th ==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>== 4th ==</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>* made cryo stocks of K1319042 and K131026 in NEB/BL21/<del class="diffchange diffchange-inline">DH5alpha</del>, I746909 in BL21 and pET17-His-SNAP-YFP-Gal3 in ''E.&nbsp;coli'' rosetta (DE3), respectively.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>* made cryo stocks of K1319042 and K131026 in NEB/BL21/<ins class="diffchange diffchange-inline">DH5α</ins>, I746909 in BL21 and pET17-His-SNAP-YFP-Gal3 in ''E.&nbsp;coli'' rosetta (DE3), respectively.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* made plasmid prep, most of them using 1.5&nbsp;mL culture medium, and eluted with 1x 50&nbsp;µL of ddH{{sub|2}}O. The resulting DNA concentrations are shown below.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* made plasmid prep, most of them using 1.5&nbsp;mL culture medium, and eluted with 1x 50&nbsp;µL of ddH{{sub|2}}O. The resulting DNA concentrations are shown below.</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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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>| I746909 BL21 #3 || 49</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>| I746909 BL21 #3 || 49</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>|-</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>|-</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>| K1319042 <del class="diffchange diffchange-inline">DH5alpha </del>|| 60</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>| K1319042 <ins class="diffchange diffchange-inline">DH5α </ins>|| 60</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>|-</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>|-</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>| K131026 <del class="diffchange diffchange-inline">DH5alpha </del>|| 150</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>| K131026 <ins class="diffchange diffchange-inline">DH5α </ins>|| 150</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>|-</div></td><td class='diff-marker'> </td><td style="background: #eee; 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>| pET17-Gal3 #1 || 30.5</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>| pET17-Gal3 #1 || 30.5</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>| I746909 BL21 #1 || 2029, 947</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>| I746909 BL21 #1 || 2029, 947</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>|-</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>|-</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>| K1319042 <del class="diffchange diffchange-inline">DH5alpha </del>|| 2029, 1780</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>| K1319042 <ins class="diffchange diffchange-inline">DH5α </ins>|| 2029, 1780</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>|-</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>|-</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>| K131026 <del class="diffchange diffchange-inline">DH5alpha </del>|| 2029, 1848</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>| K131026 <ins class="diffchange diffchange-inline">DH5α </ins>|| 2029, 1848</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>|-</div></td><td class='diff-marker'> </td><td style="background: #eee; 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>| pET17-Gal3 #1 || 3086, 923, 1262</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>| pET17-Gal3 #1 || 3086, 923, 1262</div></td></tr>
</table>StefanReinholdhttp://2014.igem.org/wiki/index.php?title=Team:Aachen/Notebook/Wetlab/August&diff=319366&oldid=prevR.hanke: /* 5th */2014-10-17T12:44:26Z<p><span class="autocomment">5th</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* made alliquots of HM, 1&nbsp;L HM + glucose + supplements and 500&nbsp;ml LB</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* made alliquots of HM, 1&nbsp;L HM + glucose + supplements and 500&nbsp;ml LB</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>* The <del class="diffchange diffchange-inline">Quickchange </del>mutagenesis PCR with a tamplate K1319000 and following primers was made. The PCR was made in two steps. The first step is PCR with both primers separatly and the second step includes PCR with two PCR products mixed with each other.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>* The <ins class="diffchange diffchange-inline">'''Quikchange''' </ins>mutagenesis PCR with a tamplate K1319000 and following primers was made. The PCR was made in two steps. The first step is PCR with both primers separatly and the second step includes PCR with two PCR products mixed with each other.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>** forward primer EYFPtoREACh1_F: AGTACAACTGGAACAGCCACAACGTCTATATC</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>** forward primer EYFPtoREACh1_F: AGTACAACTGGAACAGCCACAACGTCTATATC</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>** rewerse primer EYFPtoREACh1_R: GTTGTGGCTGTTCCAGTTGTACTCCAGCTTG</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>** rewerse primer EYFPtoREACh1_R: GTTGTGGCTGTTCCAGTTGTACTCCAGCTTG</div></td></tr>
</table>R.hankehttp://2014.igem.org/wiki/index.php?title=Team:Aachen/Notebook/Wetlab/August&diff=313984&oldid=prevVeraA at 10:25, 17 October 20142014-10-17T10:25:22Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* REACh1 gets number K1319001 and REACh2 gets number K1319002.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* REACh1 gets number K1319001 and REACh2 gets number K1319002.</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>* PCR product was restricted with DpnI 60&nbsp;min at <del class="diffchange diffchange-inline">37&nbsp;°C </del>to destroy the methylated template. Then DpnI was deaktevated at <del class="diffchange diffchange-inline">80&nbsp;°C </del>during 20&nbsp;min.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>* PCR product was restricted with DpnI 60&nbsp;min at <ins class="diffchange diffchange-inline">37°C </ins>to destroy the methylated template. Then DpnI was deaktevated at <ins class="diffchange diffchange-inline">80°C </ins>during 20&nbsp;min.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* PCR product was purifired and transformated in DH5α.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* PCR product was purifired and transformated in DH5α.</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* made precultures of NEB10βand DH5 alpha cells</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* made precultures of NEB10βand DH5 alpha cells</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* inoculation of the fermenter at 11:40, and induced the fermentation of pET17-Gal3. The fermentation is expected to run 24&nbsp;h.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* inoculation of the fermenter at 11:40, and induced the fermentation of pET17-Gal3. The fermentation is expected to run 24&nbsp;h.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>* as the first Quickchange PCR for REACh2 was not sucsessful, it was reapeted as a gradient PCR with annealing temperatire <del class="diffchange diffchange-inline">55&nbsp;°C</del>, <del class="diffchange diffchange-inline">57&nbsp;°C </del>and <del class="diffchange diffchange-inline">60&nbsp;°C</del>. Then the agarose gel with PCR product samples was made.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>* as the first Quickchange PCR for REACh2 was not sucsessful, it was reapeted as a gradient PCR with annealing temperatire <ins class="diffchange diffchange-inline">55°C</ins>, <ins class="diffchange diffchange-inline">57°C </ins>and <ins class="diffchange diffchange-inline">60°C</ins>. Then the agarose gel with PCR product samples was made.</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>== 19th ==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>== 19th ==</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* Chips with K131026 and I746909 in HM were made. Images were taken every 30 min with the Geldoc</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* Chips with K131026 and I746909 in HM were made. Images were taken every 30 min with the Geldoc</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>* A PCR of J23101.E0240, K1319000, K1319001, K1319002 was run and the product was separated on a 1,2<del class="diffchange diffchange-inline">&nbsp;</del>% agarose gel</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>* A PCR of J23101.E0240, K1319000, K1319001, K1319002 was run and the product was separated on a 1,2% agarose gel</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* A SDS gel with samples of protein purification of pET17-His-SNAP-YFP-Gal3 was made to check if the purifired samples contain the target protein - YFP-Gal3. The purification or the gel resulted in two samples with a yellow color. For the SDS we also used the two samples before and the two samples after ones with the yellow color, 6&nbsp;mL. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* A SDS gel with samples of protein purification of pET17-His-SNAP-YFP-Gal3 was made to check if the purifired samples contain the target protein - YFP-Gal3. The purification or the gel resulted in two samples with a yellow color. For the SDS we also used the two samples before and the two samples after ones with the yellow color, 6&nbsp;mL. </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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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* made chips of K131026 in NEB , DH5α and BL21 in LB and additionally K131026 in DH5α in HM+. Images were taken every 30 min with our own device</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* made chips of K131026 in NEB , DH5α and BL21 in LB and additionally K131026 in DH5α in HM+. Images were taken every 30 min with our own device</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>{{Team:Aachen/Figure|Aachen_24_08_2014_K131026_bl21_serie.png|title=Sensor Chips with K131026 in BL21 in LB taken with first prototyp of our own device|subtitle=Sensor chips with K131026 in BL21 in LB medium with 1,5<del class="diffchange diffchange-inline">&nbsp;</del>% agar, lower chip induced. A) befor induction with 2&nbsp;µl of 5000&nbsp;µg/ml HSL (3-oxo-C12) B) 0.5&nbsp;h after induction C) 1&nbsp;h after induction D) 1.5&nbsp;h after induction E) 2&nbsp;h after induction F) 2.5&nbsp;h after induction|width=900px}}</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>{{Team:Aachen/Figure|Aachen_24_08_2014_K131026_bl21_serie.png|title=Sensor Chips with K131026 in BL21 in LB taken with first prototyp of our own device|subtitle=Sensor chips with K131026 in BL21 in LB medium with 1,5% agar, lower chip induced. A) befor induction with 2&nbsp;µl of 5000&nbsp;µg/ml HSL (3-oxo-C12) B) 0.5&nbsp;h after induction C) 1&nbsp;h after induction D) 1.5&nbsp;h after induction E) 2&nbsp;h after induction F) 2.5&nbsp;h after induction|width=900px}}</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></center></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></center></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>== 25th ==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>== 25th ==</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* did a plasmid restriction of I20260 (EcoRI,PstI), J23115 (EcoRI, SpeI), K516032 (XbaI,PstI), and J23101 (EcoRI, SpeI)</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* did a plasmid restriction of I20260 (EcoRI,PstI), J23115 (EcoRI, SpeI), K516032 (XbaI,PstI), and J23101 (EcoRI, SpeI)</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>* tested the growth of ''Pseudomonas fluorescens'' in different liquid media for high OD and strong fluorescence. She tested Standard I medium, Cetrimide medium and Pseudomonas-F medium, and Pseudomonas-F medium supplemented with 300&nbsp;µL Fe3+ in 500&nbsp;mL flasks with a filling volume of 30&nbsp;mL. The flasks were inoculated with ''P. fluorescens'' cells on Standard I agar, and incubated at <del class="diffchange diffchange-inline">30&nbsp;°C </del>at 250&nbsp;rpm.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>* tested the growth of ''Pseudomonas fluorescens'' in different liquid media for high OD and strong fluorescence. She tested Standard I medium, Cetrimide medium and Pseudomonas-F medium, and Pseudomonas-F medium supplemented with 300&nbsp;µL Fe3+ in 500&nbsp;mL flasks with a filling volume of 30&nbsp;mL. The flasks were inoculated with ''P. fluorescens'' cells on Standard I agar, and incubated at <ins class="diffchange diffchange-inline">30°C </ins>at 250&nbsp;rpm.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* prepared over night cultures of K131026 in DH5α and NEB for chips</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* prepared over night cultures of K131026 in DH5α and NEB for chips</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* prepared 2x 5&nbsp;ml of pSB1C3, psB3K3 and pSB1A2 for plasmid prep</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* prepared 2x 5&nbsp;ml of pSB1C3, psB3K3 and pSB1A2 for plasmid prep</div></td></tr>
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<td colspan="2" class="diff-lineno">Line 301:</td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* restriciton of synthesized TEV protease with EcoRI and PstI</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* restriciton of synthesized TEV protease with EcoRI and PstI</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* qualitatively tested the ''Pseudomonas fluorescens'' that had grown over night for OD and fluorescence. She determined that Pseudomonas-F medium is the most adequate for the cultivation of the strain we use, since both OD and fluorescence were best in the flask containing the respective medium. Growth in the Pseudomonas-F medium supplemented with 300&nbsp;µg/L Fe3+ was weaker, however, fluorescence was also successfully suppressed.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* qualitatively tested the ''Pseudomonas fluorescens'' that had grown over night for OD and fluorescence. She determined that Pseudomonas-F medium is the most adequate for the cultivation of the strain we use, since both OD and fluorescence were best in the flask containing the respective medium. Growth in the Pseudomonas-F medium supplemented with 300&nbsp;µg/L Fe3+ was weaker, however, fluorescence was also successfully suppressed.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>* made chips with K131026 in DH5α and NEB, in LB and LB + 10<del class="diffchange diffchange-inline">&nbsp;</del>% glycerol. Images were taken with our own device every 10 min (illumination problems).</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>* made chips with K131026 in DH5α and NEB, in LB and LB + 10% glycerol. Images were taken with our own device every 10 min (illumination problems).</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>{{Team:Aachen/Figure|Aachen_26_08_2014_K131026_neb_serie.png|title=Sensor Chips with K131026 in NEB in LB taken with first prototyp of our own device|subtitle=Sensor chips with K131026 in NEB in LB medium with 1,5<del class="diffchange diffchange-inline">&nbsp;</del>% agar. Chip on the top induced with 0.5&nbsp;µl of 500&nbsp;µg/ml HSL and on the bottom with less than 0.2&nbsp;µl of 500&nbsp;µg/ml HSL (3-oxo-C12). A) befor induction B) 1&nbsp;h after induction C) 1.5&nbsp;h after induction D) 2&nbsp;h after induction |width=600px}}</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>{{Team:Aachen/Figure|Aachen_26_08_2014_K131026_neb_serie.png|title=Sensor Chips with K131026 in NEB in LB taken with first prototyp of our own device|subtitle=Sensor chips with K131026 in NEB in LB medium with 1,5% agar. Chip on the top induced with 0.5&nbsp;µl of 500&nbsp;µg/ml HSL and on the bottom with less than 0.2&nbsp;µl of 500&nbsp;µg/ml HSL (3-oxo-C12). A) befor induction B) 1&nbsp;h after induction C) 1.5&nbsp;h after induction D) 2&nbsp;h after induction |width=600px}}</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></center></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></center></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* plasmid prep of the back bones, restriction and gel purification</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* plasmid prep of the back bones, restriction and gel purification</div></td></tr>
</table>VeraAhttp://2014.igem.org/wiki/index.php?title=Team:Aachen/Notebook/Wetlab/August&diff=311439&oldid=prevVeraA: /* 5th */2014-10-17T09:15:52Z<p><span class="autocomment">5th</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 09:15, 17 October 2014</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* REACh1 gets number K1319001 and REACh2 gets number K1319002.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* REACh1 gets number K1319001 and REACh2 gets number K1319002.</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>* PCR product was restricted with DpnI <del class="diffchange diffchange-inline">20</del>&nbsp;min at 37&nbsp;°C to destroy the methylated template. Then DpnI was deaktevated at 80&nbsp;°C during 20&nbsp;min.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>* PCR product was restricted with DpnI <ins class="diffchange diffchange-inline">60</ins>&nbsp;min at 37&nbsp;°C to destroy the methylated template. Then DpnI was deaktevated at 80&nbsp;°C during 20&nbsp;min.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* PCR product was purifired and transformated in DH5α.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* PCR product was purifired and transformated in DH5α.</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>VeraAhttp://2014.igem.org/wiki/index.php?title=Team:Aachen/Notebook/Wetlab/August&diff=310906&oldid=prevVeraA: /* 5th */2014-10-17T09:00:52Z<p><span class="autocomment">5th</span></p>
<table style="background-color: white; color:black;">
<col class='diff-marker' />
<col class='diff-content' />
<col class='diff-marker' />
<col class='diff-content' />
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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 09:00, 17 October 2014</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></center></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></center></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;">* After PCR 5&nbsp;µL of each sample of PCR product were used for agarose gel, the rest was restricted with DpnI 20&nbsp;min at 37&nbsp;°C. Then DpnI was deaktevated at 80&nbsp;°C during 20&nbsp;min.</del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{Team:Aachen/Figure|Aachen_14-07-5_PCR_toREACh1%262.png|title=Agarose gel with PCR products after Quichchange from K1319000 to K1319001 and K1319002 |width=400px}}</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{Team:Aachen/Figure|Aachen_14-07-5_PCR_toREACh1%262.png|title=Agarose gel with PCR products after Quichchange from K1319000 to K1319001 and K1319002 |width=400px}}</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: #eee; color:black; font-size: smaller;"><div>* REACh1 gets number K1319001 and REACh2 gets number K1319002.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>* REACh1 gets number K1319001 and REACh2 gets number K1319002.</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;">* PCR product was restricted with DpnI 20&nbsp;min at 37&nbsp;°C to destroy the methylated template. Then DpnI was deaktevated at 80&nbsp;°C during 20&nbsp;min.</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;">* PCR product was purifired and transformated in DH5α.</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>== 6th ==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>== 6th ==</div></td></tr>
</table>VeraAhttp://2014.igem.org/wiki/index.php?title=Team:Aachen/Notebook/Wetlab/August&diff=310813&oldid=prevVeraA: /* 5th */2014-10-17T08:57:58Z<p><span class="autocomment">5th</span></p>
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<td colspan='2' style="background-color: white; color:black;">Revision as of 08:57, 17 October 2014</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></center></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></center></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> </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">* After PCR 5&nbsp;µL of each sample of PCR product were used for agarose gel, the rest was restricted with DpnI 20&nbsp;min at 37&nbsp;°C. Then DpnI was deaktevated at 80&nbsp;°C during 20&nbsp;min.</ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{Team:Aachen/Figure|Aachen_14-07-5_PCR_toREACh1%262.png|title=Agarose gel with PCR products after Quichchange from K1319000 to K1319001 and K1319002 |width=400px}}</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{Team:Aachen/Figure|Aachen_14-07-5_PCR_toREACh1%262.png|title=Agarose gel with PCR products after Quichchange from K1319000 to K1319001 and K1319002 |width=400px}}</div></td></tr>
</table>VeraAhttp://2014.igem.org/wiki/index.php?title=Team:Aachen/Notebook/Wetlab/August&diff=290025&oldid=prevVeraA: /* 6th */2014-10-16T21:38:47Z<p><span class="autocomment">6th</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 21:38, 16 October 2014</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>{{Team:Aachen/Figure|Aachen_14-08-06_to_REACh2.png|title=Agarose gel with PCR products after <del class="diffchange diffchange-inline">Quichchange </del>from K1319000 to K1319002 |width=400px}}</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>{{Team:Aachen/Figure|Aachen_14-08-06_to_REACh2.png|title=Agarose gel with PCR products after <ins class="diffchange diffchange-inline">Quickchange </ins>from K1319000 to K1319002 |width=400px}}</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></center></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></center></div></td></tr>
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</table>VeraA