Team:Brasil-SP/Notebook

From 2014.igem.org

(Difference between revisions)
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       <li>PCR lasR (BBa_C0079) for amplification + RBS addition using primers</li>
       <li>PCR lasR (BBa_C0079) for amplification + RBS addition using primers</li>
     </ul>
     </ul>
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     </th>
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     </td>
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     <th with="14,3%">01/07  
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     <td>01/07  
     <ul>
     <ul>
       <li>Purification of lasR PCR product </li>
       <li>Purification of lasR PCR product </li>
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       </ul>
       </ul>
     </ul>
     </ul>
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     </th>
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     </td>
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     <th with="14,3%">03/07
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     <td03/07
     <ul>
     <ul>
       <li>Transformation of the Biobricks sent by the Imperial College Team:</li>
       <li>Transformation of the Biobricks sent by the Imperial College Team:</li>
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       </ul>
       </ul>
     </ul>
     </ul>
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     </th>
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     </td>
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     <th with="14,3%">04/07</th>
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     <td>04/07</td>
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     <th with="14,3%">05/07</th>
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     <td>05/07</td>
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     <th with="14,2%">06/07
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     <td>06/07
     <ul>
     <ul>
       <li>Inoculum:</li>
       <li>Inoculum:</li>
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       </ul>
       </ul>
     </ul>
     </ul>
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     </th>
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     </td>
   </tr>
   </tr>
  <tr>
  <tr>
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     <th>07/07
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     <td>07/07
     <ul>
     <ul>
       <li>Miniprep, Quantification and Restriction Analysis:
       <li>Miniprep, Quantification and Restriction Analysis:
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       </ul>
       </ul>
     </ul>
     </ul>
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     </th>
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     </td>
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     <th>08/07</th>
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     <td>08/07</td>
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     <th>09/07  
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     <td>09/07  
     <ul>
     <ul>
     <li>Inoculum:</li>
     <li>Inoculum:</li>
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       </ul>
       </ul>
     </ul>
     </ul>
-
     </th>
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     </td>
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     <th>10/07
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     <td>10/07
     <ul>
     <ul>
       <li>Miniprep, Quantification and Restriction Analysis:</li>
       <li>Miniprep, Quantification and Restriction Analysis:</li>
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       <li>Ligation of RBS+lasR in PTZ57R/T vector (instaclone kit)</li>
       <li>Ligation of RBS+lasR in PTZ57R/T vector (instaclone kit)</li>
     </ul>
     </ul>
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     </th>
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     </td>
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     <th>11/07
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     <td>11/07
     <ul>
     <ul>
       <li>Prepare 40% glycerol solution</li>
       <li>Prepare 40% glycerol solution</li>
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     </ul>
     </ul>
     </th>
     </th>
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     <th>12/07</th>
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     <td>12/07</td>
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     <th>13/07</th>
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     <td>13/07</td>
   </tr> <tr>
   </tr> <tr>
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     <th>14/07
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     <td>14/07
     <ul>
     <ul>
       <li>Assembly:<a href="https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf">AIII</a></li>
       <li>Assembly:<a href="https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf">AIII</a></li>
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       <li>Prepare more LB medium (solid + liquid)</li>
       <li>Prepare more LB medium (solid + liquid)</li>
     </ul>
     </ul>
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     </th>
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     </td>
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     <th>15/07
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     <td>15/07
     <ul>
     <ul>
       <li>Assembly:<a href="https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf">AIII</a></li>
       <li>Assembly:<a href="https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf">AIII</a></li>
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       </ul>
       </ul>
     </ul>
     </ul>
-
     </th>
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     </td>
-
     <th>16/07
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     <td>16/07
     <ul>
     <ul>
     <li>Tranformation:</li>
     <li>Tranformation:</li>
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       </ul>
       </ul>
     </ul>
     </ul>
-
     </th>
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     </td>
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     <th>17/07
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     <td>17/07
     <ul>
     <ul>
       <li>Glycerol Stock and Miniprep:<li>
       <li>Glycerol Stock and Miniprep:<li>
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       </ul>
       </ul>
     </ul>
     </ul>
-
     </th>
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     </td>
-
     <th>18/07
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     <td>18/07
     <ul>
     <ul>
       <li>Glycerol Stock and Miniprep:</li>
       <li>Glycerol Stock and Miniprep:</li>
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       </ul>
       </ul>
     </ul>
     </ul>
-
     </th>
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     </td>
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     <th>19/07</th>
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     <td>19/07</th>
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     <th>20/07</th>
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     <td>20/07</th>
   </tr> <tr>
   </tr> <tr>
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     <th>21/07
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     <td>21/07
     <ul>
     <ul>
       <li>Transformation:</li>
       <li>Transformation:</li>
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       </ul>
       </ul>
     </ul>
     </ul>
-
     </th>
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     </td>
-
     <th>22/07
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     <td>22/07
     <ul>
     <ul>
       <li>Inoculum:</li>
       <li>Inoculum:</li>
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       </ul>
       </ul>
     </ul>
     </ul>
-
     </th>
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     </td>
-
     <th>23/07
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     <td>23/07
     <ul>
     <ul>
       <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf">AIII</a> and <a href="https://static.igem.org/mediawiki/2014/9/90/KI.pdf">KI</a>:</li>
       <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf">AIII</a> and <a href="https://static.igem.org/mediawiki/2014/9/90/KI.pdf">KI</a>:</li>
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       <li>Prepare RBS+lasR and RBS+qteE for sequencing</li>
       <li>Prepare RBS+lasR and RBS+qteE for sequencing</li>
     </ul>
     </ul>
-
     </th>
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     </td>
-
     <th>24/07
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     <td>24/07
     <ul>
     <ul>
       <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf">AIII</a> and <a href="https://static.igem.org/mediawiki/2014/9/90/KI.pdf">KI</a>:</li>
       <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf">AIII</a> and <a href="https://static.igem.org/mediawiki/2014/9/90/KI.pdf">KI</a>:</li>
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       </ul>
       </ul>
     </ul>
     </ul>
-
     </th>
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     </td>
-
     <th>25/07
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     <td>25/07
     <ul>
     <ul>
       <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf">AIII</a> and <a href="https://static.igem.org/mediawiki/2014/9/90/KI.pdf">KI</a>:</li>
       <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf">AIII</a> and <a href="https://static.igem.org/mediawiki/2014/9/90/KI.pdf">KI</a>:</li>
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       </ul>
       </ul>
     </ul>
     </ul>
-
     </th>
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     </td>
-
     <th>26/07</th>
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     <td>26/07</td>
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     <th>27/07</th>
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     <td>27/07</td>
   </tr> <tr>
   </tr> <tr>
-
     <th>28/07
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     <td>28/07
     <ul>
     <ul>
       <li>Assembly <a href="https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf">AIII</a>:</li>
       <li>Assembly <a href="https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf">AIII</a>:</li>
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       </ul>  
       </ul>  
     </ul>
     </ul>
-
     </th>
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     </td>
-
     <th>29/07
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     <td>29/07
     <ul>
     <ul>
       <li>>Assembly <a href="https://static.igem.org/mediawiki/2014/3/38/AII.pdf">AII</a>,
       <li>>Assembly <a href="https://static.igem.org/mediawiki/2014/3/38/AII.pdf">AII</a>,
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       </ul>
       </ul>
     </ul>
     </ul>
-
     </th>
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     </td>
-
     <th>30/07
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     <td>30/07
     <ul>
     <ul>
       <li>>Assembly <a href="https://static.igem.org/mediawiki/2014/3/38/AII.pdf">AII</a>,
       <li>>Assembly <a href="https://static.igem.org/mediawiki/2014/3/38/AII.pdf">AII</a>,
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       </ul>
       </ul>
     </ul>     
     </ul>     
-
     </th>
+
     </td>
-
     <th>31/07
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     <td>31/07
     <ul>
     <ul>
       <li>Assembly <a href="https://static.igem.org/mediawiki/2014/3/38/AII.pdf">AII</a>,
       <li>Assembly <a href="https://static.igem.org/mediawiki/2014/3/38/AII.pdf">AII</a>,
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       <li>PCR of Plac BBa_K143055 was followed by a Gel Purification and Ligation in the PUC19 vector</li>
       <li>PCR of Plac BBa_K143055 was followed by a Gel Purification and Ligation in the PUC19 vector</li>
     </ul>
     </ul>
-
     </th>
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     </td>
-
     <th></th>
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     <td></td>
-
     <th></th>
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     <td></td>
-
     <th></th>
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     <td></td>
   </tr>
   </tr>
</table>
</table>
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<table style="width:100%">
<table style="width:100%">
   <tr>
   <tr>
-
     <th>Monday</th>
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     <td>Monday</td>
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     <th>Tuesday</th>
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     <td>Tuesday</td>
-
     <th>Wednesday</th>
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     <td>Wednesday</td>
-
     <th>Thursday</th>
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     <td>Thursday</td>
-
     <th>Friday</th>
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     <td>Friday</td>
-
     <th>Saturday</th>
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     <td>Saturday</td>
-
     <th>Sunday</th>
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     <td>Sunday</td>
   </tr> <tr>
   </tr> <tr>
-
     <th></th>
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     <td></td>
-
     <th></th>
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     <td></td>
-
     <th></th>
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     <td></td>
-
     <th></th>
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     <td></td>
-
     <th>01/08
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     <td>01/08
     <ul>
     <ul>
       <li>Transformation:</li>
       <li>Transformation:</li>
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       </ul>
       </ul>
     </ul>
     </ul>
-
     </th>
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     </td>
-
     <th>02/08</th>
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     <td>02/08</td>
-
     <th>03/08</th>
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     <td>03/08</td>
   </tr>
   </tr>
  <tr>
  <tr>
-
     <th>04/08
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     <td>04/08
     <ul>
     <ul>
       <li>Inoculum</li>
       <li>Inoculum</li>
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       </ul>
       </ul>
     </ul>
     </ul>
-
     </th>
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     </td>
-
     <th>05//08
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     <td>05//08
     <ul>
     <ul>
       <li>Prepare assemblies <a href="https://static.igem.org/mediawiki/2014/9/90/KI.pdf">KI</a> for cytometry characterization</li>
       <li>Prepare assemblies <a href="https://static.igem.org/mediawiki/2014/9/90/KI.pdf">KI</a> for cytometry characterization</li>
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       </ul>
       </ul>
     </ul>
     </ul>
-
     </th>
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     </td>
-
     <th>06/08</th>
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     <td>06/08</td>
-
     <th>07/08
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     <td>07/08
     <ul>
     <ul>
       <li>MIniprep and Restriction analysis:</li>
       <li>MIniprep and Restriction analysis:</li>
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       </ul>
       </ul>
     </ul>
     </ul>
-
     </th>
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     </td>
-
     <th>08/08</th>
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     <td>08/08</td>
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     <th>09/08</th>
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     <td>09/08</td>
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     <th>10/08</th>
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     <td>10/08</td>
   </tr> <tr>
   </tr> <tr>
-
     <th>11/08
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     <td>11/08
     <ul>
     <ul>
       <li>Prepare Plac BBa_K143055 for sequencing</li>
       <li>Prepare Plac BBa_K143055 for sequencing</li>
       <li>Analyse the RBS+qteE and RBS+lasR sequencing file</li>
       <li>Analyse the RBS+qteE and RBS+lasR sequencing file</li>
     </ul>
     </ul>
-
     </th>
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     </td>
-
     <th>12/08
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     <td>12/08
     <ul>
     <ul>
       <li>Digestion:</li>
       <li>Digestion:</li>
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       </ul>
       </ul>
     </ul>
     </ul>
-
     </th>
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     </td>
-
     <th>13/08
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     <td>13/08
     <ul>
     <ul>
       <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/4/48/BII.pdf">BII</a> and  
       <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/4/48/BII.pdf">BII</a> and  
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       </ul>
       </ul>
     </ul>
     </ul>
-
     </th>
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     </td>
-
     <th>14/08
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     <td>14/08
     <ul>
     <ul>
       <li>Gel Purification and Ligation in pSB1C3 for BBiobrick standard:</li>
       <li>Gel Purification and Ligation in pSB1C3 for BBiobrick standard:</li>
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       <li>OBS: we had some problems here with the purification and we lost all our digestions, so we're repeating the digestion</li>
       <li>OBS: we had some problems here with the purification and we lost all our digestions, so we're repeating the digestion</li>
     </ul>
     </ul>
-
     </th>
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     </td>
-
     <th>15/08</th>
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     <td>15/08</td>
-
     <th>16/08
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     <td>16/08
     <ul>
     <ul>
       <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/4/48/BII.pdf">BII</a> and  
       <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/4/48/BII.pdf">BII</a> and  
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       </ul>
       </ul>
     </ul>
     </ul>
-
     </th>
+
     </td>
-
     <th>17/08
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     <td>17/08
     <ul>
     <ul>
       <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/4/48/BII.pdf">BII</a> and  
       <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/4/48/BII.pdf">BII</a> and  
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       </ul>
       </ul>
     </ul>
     </ul>
-
     </th>
+
     </td>
   </tr> <tr>
   </tr> <tr>
-
     <th>18/08
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     <td>18/08
     <ul>
     <ul>
       <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/4/48/BII.pdf">BII</a> and  
       <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/4/48/BII.pdf">BII</a> and  
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       <li>Prepare LB medium (liquid and solid)</li>
       <li>Prepare LB medium (liquid and solid)</li>
     </ul>
     </ul>
-
     </th>
+
     </td>
-
     <th>19/08
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     <td>19/08
     <ul>
     <ul>
       <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/4/48/BII.pdf">BII</a> and  
       <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/4/48/BII.pdf">BII</a> and  
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       </ul>
       </ul>
     </ul>
     </ul>
-
     </th>
+
     </td>
-
     <th>20/08
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     <td>20/08
     <ul>
     <ul>
       <li>RBS+qteE, RBS+lasR and Assemblies <a href="https://static.igem.org/mediawiki/2014/4/48/BII.pdf">BII</a> and  
       <li>RBS+qteE, RBS+lasR and Assemblies <a href="https://static.igem.org/mediawiki/2014/4/48/BII.pdf">BII</a> and  
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       </ul>
       </ul>
     </ul>
     </ul>
-
     </th>
+
     </td>
-
     <th>21/08
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     <td>21/08
     <ul>
     <ul>
       <li>RBS+qteE, RBS+lasR and Assemblies <a href="https://static.igem.org/mediawiki/2014/4/48/BII.pdf">BII</a> and  
       <li>RBS+qteE, RBS+lasR and Assemblies <a href="https://static.igem.org/mediawiki/2014/4/48/BII.pdf">BII</a> and  
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       </ul>
       </ul>
     </ul>
     </ul>
-
     </th>
+
     </td>
-
     <th>22/08
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     <td>22/08
     <ul>
     <ul>
       <li>Prepare LB medium (liquid and solid)</li>
       <li>Prepare LB medium (liquid and solid)</li>
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       </ul>
       </ul>
     </ul>
     </ul>
-
     </th>
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     </td>
-
     <th>23/08</th>
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     <td>23/08</td>
-
     <th>24/08</th>
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     <td>24/08</td>
   </tr> <tr>
   </tr> <tr>
-
     <th>25/08
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     <td>25/08
     <ul>
     <ul>
       <li>Inoculum</li>
       <li>Inoculum</li>
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       <li>PCR BBa_K143055 with higher melting temperature (check out our Lab Journal for more info)</li>
       <li>PCR BBa_K143055 with higher melting temperature (check out our Lab Journal for more info)</li>
     <ul>
     <ul>
-
     </th>
+
     </td>
-
     <th>26/08
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     <td>26/08
     <ul>
     <ul>
       <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/b/b7/AI.pdf">AI</a>,  
       <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/b/b7/AI.pdf">AI</a>,  
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       </ul>
       </ul>
     </ul>
     </ul>
-
     </th>
+
     </td>
-
     <th>27/08
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     <td>27/08
     <ul>
     <ul>
       <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/b/b7/AI.pdf">AI</a>,  
       <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/b/b7/AI.pdf">AI</a>,  
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       </ul>
       </ul>
     </ul>
     </ul>
-
     </th>
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     </td>
-
     <th>28/08</th>
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     <td>28/08</td>
-
     <th>29/08</th>
+
     <td>29/08</td>
-
     <th>30/08</th>
+
     <td>30/08</td>
-
     <th>31/08</th>
+
     <td>31/08</td>
   </tr>
   </tr>
</table>
</table>
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<table style="width:100%">
<table style="width:100%">
   <tr>
   <tr>
-
     <th>Monday</th>
+
     <td>Monday</td>
-
     <th>Tuesday</th>
+
     <td>Tuesday</td>
-
     <th>Wednesday</th>
+
     <td>Wednesday</td>
-
     <th>Thursday</th>
+
     <td>Thursday</td>
-
     <th>Friday</th>
+
     <td>Friday</td>
-
     <th>Saturday</th>
+
     <td>Saturday</td>
-
     <th>Sunday</th>
+
     <td>Sunday</td>
   </tr> <tr>
   </tr> <tr>
-
     <th>01/09
+
     <td>01/09
     <ul>
     <ul>
       <li>Assembly <a href="https://static.igem.org/mediawiki/2014/a/a3/CII.pdf">CII</a>
       <li>Assembly <a href="https://static.igem.org/mediawiki/2014/a/a3/CII.pdf">CII</a>
Line 816: Line 816:
       </ul>
       </ul>
     </ul>
     </ul>
-
     </th>
+
     </td>
-
     <th>02/09
+
     <td>02/09
     <ul>
     <ul>
       <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/b/b7/AI.pdf">AI</a>,  
       <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/b/b7/AI.pdf">AI</a>,  
Line 828: Line 828:
       </ul>
       </ul>
     </ul>
     </ul>
-
     </th>
+
     </td>
-
     <th>03/09
+
     <td>03/09
     <ul>
     <ul>
       <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/b/b7/AI.pdf">AI</a>,  
       <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/b/b7/AI.pdf">AI</a>,  
Line 840: Line 840:
       </ul>
       </ul>
     </ul>
     </ul>
-
     </th>
+
     </td>
-
     <th>04/09
+
     <td>04/09
     <ul>
     <ul>
       <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/b/b7/AI.pdf">AI</a>,  
       <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/b/b7/AI.pdf">AI</a>,  
Line 854: Line 854:
       </ul>
       </ul>
     </ul>
     </ul>
-
     </th>
+
     </td>
-
     <th>05/09</th>
+
     <td>05/09</td>
-
     <th>06/09</th>
+
     <td>06/09</td>
-
     <th>07/09</th>
+
     <td>07/09</td>
   </tr>
   </tr>
  <tr>
  <tr>
-
     <th>08/09
+
     <td>08/09
     <ul>
     <ul>
       <li>QuickChange PCR to mutate the cleavage site of the BBa_316037 to a Cathepsin S recognition site</li>
       <li>QuickChange PCR to mutate the cleavage site of the BBa_316037 to a Cathepsin S recognition site</li>
       <li>Transformation of the QuickCHange PCR product</li>
       <li>Transformation of the QuickCHange PCR product</li>
     </ul>
     </ul>
-
     </th>
+
     </td>
-
     <th>09/09</th>
+
     <td>09/09</td>
-
     <th>10/09
+
     <td>10/09
     <ul>
     <ul>
       <li>The QuickChange protocol didn't work well. So try again with more attention!</li>
       <li>The QuickChange protocol didn't work well. So try again with more attention!</li>
Line 880: Line 880:
       </ul>  
       </ul>  
     </ul>
     </ul>
-
     </th>
+
     </td>
-
     <th>11/09
+
     <td>11/09
     <ul>
     <ul>
       <li>Prepare inoculum of the mutated BBa_K316037</li>
       <li>Prepare inoculum of the mutated BBa_K316037</li>
Line 893: Line 893:
       </ul>   
       </ul>   
     </ul>
     </ul>
-
     </th>
+
     </td>
-
     <th>12/09
+
     <td>12/09
     <ul>
     <ul>
       <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/7/72/DI.pdf">DI</a>,  
       <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/7/72/DI.pdf">DI</a>,  
Line 906: Line 906:
       <li>PCR for BBa_143015 (BBa_143055 without RBS)</li>
       <li>PCR for BBa_143015 (BBa_143055 without RBS)</li>
     </ul>
     </ul>
-
     </th>
+
     </td>
-
     <th>13/09
+
     <td>13/09
     <ul>
     <ul>
       <li>Assembly <a href="https://static.igem.org/mediawiki/2014/9/95/KX.pdf">KX</a>  
       <li>Assembly <a href="https://static.igem.org/mediawiki/2014/9/95/KX.pdf">KX</a>  
Line 921: Line 921:
       <li>Prepare LB medium (solid and liquid)</li>
       <li>Prepare LB medium (solid and liquid)</li>
     </ul>
     </ul>
-
     </th>
+
     </td>
-
     <th>14/09
+
     <td>14/09
     <ul>
     <ul>
       <li>Assembly <a href="https://static.igem.org/mediawiki/2014/9/95/KX.pdf">KX</a>  
       <li>Assembly <a href="https://static.igem.org/mediawiki/2014/9/95/KX.pdf">KX</a>  
Line 936: Line 936:
       </ul>
       </ul>
     </ul>
     </ul>
-
     </th>
+
     </td>
   </tr> <tr>
   </tr> <tr>
-
     <th>15/09
+
     <td>15/09
     <ul>
     <ul>
       <li>Assembly <a href="https://static.igem.org/mediawiki/2014/9/95/KX.pdf">KX</a>  
       <li>Assembly <a href="https://static.igem.org/mediawiki/2014/9/95/KX.pdf">KX</a>  
Line 952: Line 952:
       <li>Gel Purification of the BBa_K143015</li>
       <li>Gel Purification of the BBa_K143015</li>
     </ul>
     </ul>
-
     </th>
+
     </td>
-
     <th>16/09
+
     <td>16/09
     <ul>
     <ul>
       <li>Prepare 40% glycerol solution</li>
       <li>Prepare 40% glycerol solution</li>
Line 965: Line 965:
       </ul>
       </ul>
     </ul>
     </ul>
-
     </th>
+
     </td>
-
     <th>17/09
+
     <td>17/09
     <ul>
     <ul>
       <li>LIgation of the synthesized BBa_K143015 in TOPO vector followed by Transformation</li>
       <li>LIgation of the synthesized BBa_K143015 in TOPO vector followed by Transformation</li>
     </ul>
     </ul>
-
     </th>
+
     </td>
-
     <th>18/09
+
     <td>18/09
     <ul>
     <ul>
       <li>Replate BBa_K143015. We forgot the X-gal :(</li>
       <li>Replate BBa_K143015. We forgot the X-gal :(</li>
     </ul>
     </ul>
-
     </th>
+
     </td>
-
     <th>19/09</th>
+
     <td>19/09</td>
-
     <th>20/09</th>
+
     <td>20/09</td>
-
     <th>21/09</th>
+
     <td>21/09</td>
   </tr> <tr>
   </tr> <tr>
-
     <th>22/09
+
     <td>22/09
     <ul>
     <ul>
       <li>Inoculum of the BBa_K143015</li>
       <li>Inoculum of the BBa_K143015</li>
       <li>Assembly KXIV
       <li>Assembly KXIV
     </ul>
     </ul>
-
     </th>
+
     </td>
-
     <th>23/09</th>
+
     <td>23/09</td>
-
     <th>24/09</th>
+
     <td>24/09</td>
-
     <th>25/09</th>
+
     <td>25/09</td>
-
     <th>26/09</th>
+
     <td>26/09</td>
-
     <th>27/09</th>
+
     <td>27/09</td>
-
     <th>28/09</th>
+
     <td>28/09</td>
   </tr> <tr>
   </tr> <tr>
-
     <th>29/09</th>
+
     <td>29/09</td>
-
     <th>30/09</th>
+
     <td>30/09</td>
-
     <th></th>
+
     <td></td>
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     <th></th>
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     <th></th>
+
     <td></td>
-
     <th></th>
+
     <td></td>
   </tr>
   </tr>
</table>
</table>
<head><th>October</th>
<head><th>October</th>
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<table style="width:100%">
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   <tr>
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     <th>Monday</th>
+
     <td>Monday</td>
-
     <th>Tuesday</th>
+
     <td>Tuesday</td>
-
     <th>Wednesday</th>
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     <td>Wednesday</td>
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     <th>Thursday</th>
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     <td>Thursday</td>
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     <th>Friday</th>
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     <td>Saturday</td>
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     <th>Sunday</th>
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     <td>Sunday</td>
   </tr> <tr>
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     <th></th>
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     <td></td>
     <th>30/07</th>
     <th>30/07</th>
     <th>01/08</th>
     <th>01/08</th>

Revision as of 23:58, 27 September 2014



WELCOME TO iGEM 2014!

Your team has been approved and you are ready to start the iGEM season!
On this page you can document your project, introduce your team members, document your progress
and share your iGEM experience with the rest of the world!


Click here to edit this page!

Home Team Official Team Profile Project Parts Modeling Notebook Safety Attributions

Notebook

You should make use of the calendar feature on the wiki and start a lab notebook. This may be looked at by the judges to see how your work progressed throughout the summer. It is a very useful organizational tool as well.

Lab Protocols

Competent Cell with Calcium Chloride

  1. Materials

    • Solid LB medium - 1 plate
    • Liquid LB medium
    • 2 250ml centrifuge tubes
    • Centrifuge
    • Cell sample (DH5-alpha in our case)
    • Autoclave
    • Inoculation loop
    • Liquid nitrogen
    • 37°C oven
    • CaCl2:
      • 60mM CaCl2
      • 10mM HEPES
      • 15% glicerol
      • H2O to 100ml

  2. Methods

    3. Day 1

    1. Risk the LB plate with the cell sample (DH5-alpha)
    2. Incubate the plate at 37°C overnight

    Day 2

    1. Sterilize in the Autoclave
      • 100ml of liquid LB medium
      • 100 ml of CaCl2
      • 2 250ml centrifuge tubes
    2. Prepare a 6ml LB inoculum with a DH5-alpha colony and incubate at 37°C/250rpm overnight

    Day 3

    1. Add 2ml of the inoculum in 100ml of liquid LB medium. Incubate at 37°C/250rpm until the OD reaches 0,375
    2. Distribute the volume in 2 250ml centrifuge tubes (pre chilled) and spin at 10000rpm/7min/4°C
    3. Discard the supernatant and ressuspend the pellet in 10ml of CaCl2 solution
    4. Spin the tubes at 7000rpm/7min/4°C
    5. Discard the supernatant and ressuspend the pellet in 10ml of CaCl2 solution. Incubate 30min on ice
    6. Spin at 7000rpm/7min/4°C
    7. Discard the supernatant and ressuspend the pellet in 2ml of the CaCl2 solution
    8. Distribute the 2ml in 500ul tubes adding 50ul in each.
    9. Freeze the samples in liquid nitrogen
    10. Stock at -80°C

Transformation in Escherichia coli (DH5-alpha)

  1. Materials

    • 1,5ml tube
    • Styrofoam box with ice
    • Agar plate with antibiotic
    • Competent cells
    • Plasmidial DNA
    • Centrifuge
    • Water bath at 42°C
    • Liquid LB medium
    • Shaker

  2. Methods

    1. Briefly spin the competent cells and put then on ice
    2. Add 50ng of plasmidial DNA in a 1,5ml tube
    3. Add the 50ul of competent cell in the same tube
    4. Keep the tube on ice for 25min
    5. Put the tube in a 42° water bath for 2 min
    6. Put the tube on ice for 5 min
    7. Add 200ul of liquid LB
    8. Incubate at 250rpm/37°C/1 hour
    9. Plate the the solution in a Agar plate with the appropriate antibiotic
    10. Incubate the plate at 37°C overnight

Assemblies

Life Inside the LAB

July
  • Transformation of the Biobricks sent by the Imperial College Team:
    • BBa_K316016
    • BBa_K143031
  • Miniprep, Quantification of the plasmidial DNA samples and Restriction Analysis:
    • BBa_143012
    • BBa_K081001
    • BBa_E0840
    Monday Tuesday
      		 Wednesday Thursday Friday Saturday Sunday
      30/06
      • PCR lasR (BBa_C0079) for amplification + RBS addition using primers
      		 01/07
      • Purification of lasR PCR product
      		02/07
      • Transformation of the Biobricks:
        • BBa_143012
        • BBa_K081001
        • BBa_E0840
      • Inoculum of the Biobricks sent by iGEM HQ:
        • BBa_K316037
        • BBa_K316018
        • BBa_K316015
      		 04/07 05/07 06/07
      • Inoculum:
        • BBa_143012
        • BBa_K081001
        • BBa_E0840
      07/07
      • Miniprep, Quantification and Restriction Analysis:
        • BBa_143012
        • BBa_K081001
        • BBa_E0840
      • Transformation:
        • BBa_B0015
      		 08/07 09/07
      • Inoculum:
        • BBa_B0015
      		 10/07
      • Miniprep, Quantification and Restriction Analysis:
        • BBa_B0015 (restriction analysis fail, repeat)
      • PCR qteE for amplification + RBS addition using primers
      • Ligation of RBS+lasR in PTZ57R/T vector (instaclone kit)
      		 11/07
      • Prepare 40% glycerol solution
      • Transformation:
        • RBS + lasR (BBa_C0079)
      • Purification:
        • RBS + qteE
      		12/07 13/07
      14/07
      • Assembly:AIII
        • Digestion EXSP
      • Prepare X-gal
      • Prepare more LB medium (solid + liquid)
      		 15/07
      • Assembly:AIII
        • Quantification of digestion EXSP products
      • Replate RBS+lasR
      • Ligation of PCR qteE in pGEM T-easy vector
      • Transformation:
        • BBa_J04450
      • Restriction analysis:
        • BBa_B0015
      		 16/07
      • Tranformation:
        • BBa_K823003
        • RBS+qteE
      • Electrophoresis analysis:
        • BBa_B0015
      		 17/07
      • Glycerol Stock and Miniprep:
        • BBa_J04450
        • RBS+lasR
        • BBa_B0015
      • Digestion:
        • BBa_J04450 (EP)
        • BBa_J04450 (XS)
      • Inoculum:
        • BBa_K823003
      • Transformation
        • BBa_K143055
      		 18/07
      • Glycerol Stock and Miniprep:
        • BBa_J04450
        • BBa_B0015
      • Gel Purification:
        • BBa_J04450 (EP)
        • BBa_J04450 (XS)
      • Restriction analysis:
        • BBa_K823003
      		 19/07 20/07
      21/07
      • Transformation:
        • BBa_316016
      • Inoculum:
        • BBa_K143055
        • RBS+qteE
      • Assembly KI:
        • Digestion EXSP
      		 22/07
      • Inoculum:
        • BBa_K316016
      • Miniprep, Restriction analysis and Digestion:
        • RBS+qteE
      • Assemblies AIII and KI:
        • Gel Purification
        • Ligation
      		 23/07
      • Assemblies AIII and KI:
        • Transformation
      • Prepare more competent cells
      • Prepare LB medium (solid and liquid)
      • Inoculum
        • BBa_K316016
      • Prepare RBS+lasR and RBS+qteE for sequencing
      		 24/07
      • Assemblies AIII and KI:
        • Inoculum
      • Miniprep:
        • BBa_K316016
      		 25/07
      • Assemblies AIII and KI:
        • Glycerol Stock
        • Miniprep
        • Restriction analysis
      		 26/07 27/07
      28/07
      • Assembly AIII:
        • Restriction analysis (+NdeI enzyme)
        • OBS: This analysis was reapeted because we had problems confirming the assembly. The issue was that bot the construction and the pSB1C3 vector had similar size. Check out the Lab Journal report of the day for more information.
      • Assembly AII, AV and AVI:
        • Digestion EXSP
      		 29/07
      • >Assembly AII, AV and AVI:
        • Gel Purification
        • Ligation
      		 30/07
      • >Assembly AII, AV and AVI:
        • Transformation
      		 31/07
      • Assembly AII, AV and AVI:
        • Inoculum
      • PCR BBa_K143055 for RBS removal
      • OBS: This was one of the parts sent by the Imperial College Team. Because of a comunication failure we assumed that this part was comming without the RBS, so we included a RBS in the parts we were going to use with this Lac promotor :)
      • PCR of Plac BBa_K143055 was followed by a Gel Purification and Ligation in the PUC19 vector
      August
      Monday Tuesday Wednesday Thursday Friday Saturday Sunday
      01/08
      • Transformation:
        • BBa_K143055
      • Assembly AII, AV and AVI:
        • Glycerol Stock
        • Miniprep
        • Restriction analysis
      		 02/08 03/08
      04/08
      • Inoculum
        • E. coli DH5-alpha
        • KI
        • Plac BBa_K143055
      • Transformation
        • BBa_P0312
      • Assembly AV:
        • Prepare more inoculum
        • Repeat Restriction analysis (for more info go to the day's report)
      		 05//08
      • Prepare assemblies KI for cytometry characterization
      • Miniprep:
        • Plac BBa_K143055
      • Inoculum
        • BBa_P0312
      • Assembly AV:
        • Miniprep
        • And once again! Restriction analysis (this BBa_K316016 is tough)
      		 06/08 07/08
      • MIniprep and Restriction analysis:
        • BBa_P0312
      		 08/08 09/08 10/08
      11/08
      • Prepare Plac BBa_K143055 for sequencing
      • Analyse the RBS+qteE and RBS+lasR sequencing file
      		 12/08
      • Digestion:
        • RBS+qteE
        • RBS+lasR
        • OBS: this digestion was performed so we clone this parts in the pSB1C3 vector to put them in Biobrick standard
      		 13/08
      • Assemblies BII and BIII:
        • Digestion EXSP
      		 14/08
      • Gel Purification and Ligation in pSB1C3 for BBiobrick standard:
        • RBS+qteE
        • RBS+lasR
      • Assemblies BII and BIII:
      • Gel Purification
      • OBS: we had some problems here with the purification and we lost all our digestions, so we're repeating the digestion
      		 15/08 16/08
      • Assemblies BII and BIII:
        • Digestion EXSP
      		 17/08
      • Assemblies BII and BIII:
        • Gel Purification
      18/08
      • Assemblies BII and BIII:
        • Ligation
      • Prepare LB medium (liquid and solid)
      		 19/08
      • Assemblies BII and BIII:
        • Transformation
      		 20/08
      • RBS+qteE, RBS+lasR and Assemblies BII and BIII:
        • Inoculum
      		 21/08
      • RBS+qteE, RBS+lasR and Assemblies BII and BIII:
        • Glycerol Stock
        • Miniprep
        • Restriction analysis
      		 22/08
      • Prepare LB medium (liquid and solid)
      • Assembly BII:
        • Repeat restriction analysis adding the enzyme NdeI (for more info check out the Lab Journal)
      		 23/08 24/08
      25/08
      • Inoculum
        • BBa_J04450
        • BBa_K143055
      • PCR BBa_K143055 with higher melting temperature (check out our Lab Journal for more info)
      		 26/08 27/08 28/08 29/08 30/08 31/08
      September
      Monday Tuesday Wednesday Thursday Friday Saturday Sunday
      01/09
      • Assembly CII
        • Digestion of the part BII with EcoRV (for more info check out our Lab Journal)
      • Assemblies AI, CII, KIII, KIV and KVI:
        • Ligation
      		 02/09 03/09 04/09
      • Assemblies AI, CII, KIII, KIV and KVI:
        • Glycerol Stock
        • Miniprep
        • Restriction analysis
      		 05/09 06/09 07/09
      08/09
      • QuickChange PCR to mutate the cleavage site of the BBa_316037 to a Cathepsin S recognition site
      • Transformation of the QuickCHange PCR product
      		 09/09 10/09
      • The QuickChange protocol didn't work well. So try again with more attention!
      • QuickChange PCR to mutate the cleavage site of the BBa_316037 to a Cathepsin S recognition site
      • Transformation of the QuickCHange PCR product
      • Assemblies DI, KVIII, KX and KXI:
        • Digestion EXSP
      		 11/09
      • Prepare inoculum of the mutated BBa_K316037
      • Assemblies DI, KVIII, KX and KXI:
        • Gel Purification
        • Ligation
      		 12/09
      • Assemblies DI, KVIII, KX and KXI:
        • Tranformation
      • We had far to many difficulties trying to remove the RBS from the BBa_K143055, so we decided to synthesize it using PCR.
      • PCR for BBa_143015 (BBa_143055 without RBS)
      		 13/09
      • Assembly KX
        • Inoculum
      • Assemblies DI, KVIII and KXI:
        • Repeat Ligation
      • Prepare LB medium (solid and liquid)
      		 14/09
      • Assembly KX
        • Glycerol Stock
        • Miniprep
      • Assemblies DI, KVIII and KXI:
        • Transformation
      15/09
      • Assembly KX
        • Restriction analysis
      • Assemblies DI, KVIII and KXI:
        • Inoculum
      • Gel Purification of the BBa_K143015
      		 16/09
      • Prepare 40% glycerol solution
      • Assemblies DI, KVIII and KXI:
        • Glycerol Stock
        • Miniprep
        • Restriction analysis
      		 17/09
      • LIgation of the synthesized BBa_K143015 in TOPO vector followed by Transformation
      		 18/09
      • Replate BBa_K143015. We forgot the X-gal :(
      		 19/09 20/09 21/09
      22/09
      • Inoculum of the BBa_K143015
      • Assembly KXIV
      		23/09 24/09 25/09 26/09 27/09 28/09
      29/09 30/09
      October
      Monday Tuesday Wednesday Thursday Friday Saturday Sunday
      30/07 01/08 02/08 03/08 Points Points
      First Name Last Name Points Points Points Points Points
      First Name Last Name Points Points Points Points Points
      First Name Last Name Points Points Points Points Points
      First Name Last Name Points Points Points Points Points

      Retrieved from "http://2014.igem.org/Team:Brasil-SP/Notebook"