Team:Brasil-SP/Notebook

From 2014.igem.org

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<h1 align=center>Main Assembly Map</h1>
<h1 align=center>Main Assembly Map</h1>
 +
 +
 +
<p><div align="justify">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;The main assembly map describes all the constructions that would be done in the project, but some of them have not been performed in time of the iGEM deadline. The succesfully constructed assemblies is indicated by a green check point and the unsuccessfull constructions are indicated by a red "X". Click on these assemblies to access the lab form.</div></p>
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   <div class="map_image" style="background-image: url('https://static.igem.org/mediawiki/2014/7/78/Mapa_assembly_protocols.png');">
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   <div class="map_image" style="background-image: url('https://static.igem.org/mediawiki/2014/8/87/Mapa.png');">
<a class="map_link" id="map_link_1" title="" href="https://static.igem.org/mediawiki/2014/b/b7/AI.pdf">AI</a>
<a class="map_link" id="map_link_1" title="" href="https://static.igem.org/mediawiki/2014/b/b7/AI.pdf">AI</a>
<a class="map_link" id="map_link_2" title="" href="https://static.igem.org/mediawiki/2014/3/38/AII.pdf">AII</a>
<a class="map_link" id="map_link_2" title="" href="https://static.igem.org/mediawiki/2014/3/38/AII.pdf">AII</a>
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<a class="map_link" id="map_link_9" title="" href="https://static.igem.org/mediawiki/2014/d/d6/AVI.pdf">AVI</a>
<a class="map_link" id="map_link_9" title="" href="https://static.igem.org/mediawiki/2014/d/d6/AVI.pdf">AVI</a>
<a class="map_link" id="map_link_10" title="" href="https://static.igem.org/mediawiki/2014/f/fe/AV.pdf">AV</a>
<a class="map_link" id="map_link_10" title="" href="https://static.igem.org/mediawiki/2014/f/fe/AV.pdf">AV</a>
 +
        <a class="map_link" id="map_link_11" title="" href="https://static.igem.org/mediawiki/2014/5/5b/BIV.pdf">BIV</a>
   </div>
   </div>
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<h1 align=center>Assemblies forms</h1>
<h1 align=center>Assemblies forms</h1>
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         <li><a href"http://parts.igem.org/Part:BBa_B0015" style="color:#3ab473">BBa_B0015</a> (restriction analysis fail, repeat)</li>
         <li><a href"http://parts.igem.org/Part:BBa_B0015" style="color:#3ab473">BBa_B0015</a> (restriction analysis fail, repeat)</li>
       </ul>
       </ul>
-
       <li><a href="https://static.igem.org/mediawiki/2014/7/74/PCR.pdf" style="color:#f05151">PCR</a> the qteE gene for amplification. We also added the RBS (<a href="http://parts.igem.org/Part:BBa_K143021" style="color:#3ab473">BBa_K143021,</a>) using the foward primer. In our primers we only added the restriction sites X and S, so we still need to put it in the pSB1C3 for the Biobrick standard completion. When standardized this biobrick will be called BBa_K1521000.</li>
+
       <li><a href="https://static.igem.org/mediawiki/2014/7/74/PCR.pdf" style="color:#f05151">PCR</a> the qteE gene for amplification. We also added the RBS (<a href="http://parts.igem.org/Part:BBa_K143021" style="color:#3ab473">BBa_K143021,</a>) using the foward primer. In our primers we only added the restriction sites X and S, so we still need to put it in the pSB1C3 for the Biobrick standard completion. When standardized this biobrick will be called <a href="http://parts.igem.org/Part:BBa_K1521000" style="color:#3ab473">BBa_K1521000</a>.</li>
       <li><a href="https://static.igem.org/mediawiki/2014/0/03/PCR_Product_Clonig.pdf" style="color:#f05151">Ligation</a> of RBS+lasR in PTZ57R/T vector (instaclone kit)</li>
       <li><a href="https://static.igem.org/mediawiki/2014/0/03/PCR_Product_Clonig.pdf" style="color:#f05151">Ligation</a> of RBS+lasR in PTZ57R/T vector (instaclone kit)</li>
     </ul>
     </ul>
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       <li><a href="https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf" style="color:#f05151"> Transformation</a>:</li>
       <li><a href="https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf" style="color:#f05151"> Transformation</a>:</li>
       <ul>
       <ul>
-
         <li>RBS + lasR (BBa_C0079)</li>
+
         <li>RBS + lasR (<a href="http://parts.igem.org/Part:BBa_C0079"style="color:#3ab473">BBa_C0079</a>)</li>
       </ul>
       </ul>
       <li><a href="https://static.igem.org/mediawiki/2014/a/af/Agarose_Gel_Electrophoresis_and_DNA_Gel_Purification.pdf"  style="color:#f05151">Gel Purification</a>:</li>
       <li><a href="https://static.igem.org/mediawiki/2014/a/af/Agarose_Gel_Electrophoresis_and_DNA_Gel_Purification.pdf"  style="color:#f05151">Gel Purification</a>:</li>
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       <li><a href="https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf" style="color:#f05151"> Transformation</a>:</li>
       <li><a href="https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf" style="color:#f05151"> Transformation</a>:</li>
       <ul>
       <ul>
-
         <li>BBa_J04450</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_J04450" style="color:#3ab473"> <a href="" style="color:#3ab473"> BBa_J04450</a></a></li>
       </ul>
       </ul>
       <li><a href="https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf" style="color:#f05151">Restriction analysis</a>:</li>
       <li><a href="https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf" style="color:#f05151">Restriction analysis</a>:</li>
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     <li><a href="https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf" style="color:#f05151">Transformation</a>:</li>
     <li><a href="https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf" style="color:#f05151">Transformation</a>:</li>
       <ul>
       <ul>
-
       <li>BBa_K823003</li>
+
       <li><a href="http://parts.igem.org/Part:BBa_K823003" style="color:#3ab473">BBa_K823003</a></li>
       <li>RBS+qteE</li>
       <li>RBS+qteE</li>
       </ul>
       </ul>
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       <li>Glycerol Stock and <a href="https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf"style="color:#f05151">Miniprep</a>:<li>
       <li>Glycerol Stock and <a href="https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf"style="color:#f05151">Miniprep</a>:<li>
       <ul>
       <ul>
-
         <li>BBa_J04450</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_J04450" style="color:#3ab473"> BBa_J04450</a></li>
         <li>RBS+lasR</li>
         <li>RBS+lasR</li>
         <li><a href"http://parts.igem.org/Part:BBa_B0015" style="color:#3ab473">BBa_B0015</a></li>
         <li><a href"http://parts.igem.org/Part:BBa_B0015" style="color:#3ab473">BBa_B0015</a></li>
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       <li><a href="https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf" style="color:#f05151">Digestion</a>:</li>
       <li><a href="https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf" style="color:#f05151">Digestion</a>:</li>
       <ul>
       <ul>
-
         <li>BBa_J04450 (EP)</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_J04450" style="color:#3ab473"> BBa_J04450</a> (EP)</li>
-
         <li>BBa_J04450 (XS)</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_J04450" style="color:#3ab473"> BBa_J04450</a> (XS)</li>
       </ul>
       </ul>
       <li>Inoculum:</li>
       <li>Inoculum:</li>
       <ul>
       <ul>
-
         <li>BBa_K823003</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_K823003" style="color:#3ab473">BBa_K823003</a></li>
       </ul>
       </ul>
       <li><a href="https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf" style="color:#f05151"> Transformation</a></li>
       <li><a href="https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf" style="color:#f05151"> Transformation</a></li>
       <ul>
       <ul>
-
         <li>BBa_K143055</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_K143055" style="color:#3ab473">BBa_K143055</a></li>
       </ul>
       </ul>
     </ul>
     </ul>
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       <li>Glycerol Stock and <a href="https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf"style="color:#f05151">Miniprep</a>:</li>
       <li>Glycerol Stock and <a href="https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf"style="color:#f05151">Miniprep</a>:</li>
       <ul>
       <ul>
-
         <li>BBa_J04450</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_J04450" style="color:#3ab473"> BBa_J04450</a></li>
         <li><a href"http://parts.igem.org/Part:BBa_B0015" style="color:#3ab473">BBa_B0015</a></li>
         <li><a href"http://parts.igem.org/Part:BBa_B0015" style="color:#3ab473">BBa_B0015</a></li>
       </ul>
       </ul>
       <li><a href="https://static.igem.org/mediawiki/2014/a/af/Agarose_Gel_Electrophoresis_and_DNA_Gel_Purification.pdf"  style="color:#f05151">Gel Purification</a>:</li>
       <li><a href="https://static.igem.org/mediawiki/2014/a/af/Agarose_Gel_Electrophoresis_and_DNA_Gel_Purification.pdf"  style="color:#f05151">Gel Purification</a>:</li>
       <ul>
       <ul>
-
         <li>BBa_J04450 (EP)</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_J04450" style="color:#3ab473"> BBa_J04450</a> (EP)</li>
-
         <li>BBa_J04450 (XS)</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_J04450" style="color:#3ab473"> BBa_J04450</a> (XS)</li>
       </ul>
       </ul>
       <li><a href="https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf" style="color:#f05151">Restriction analysis</a>:</li>
       <li><a href="https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf" style="color:#f05151">Restriction analysis</a>:</li>
       <ul>
       <ul>
-
         <li>BBa_K823003</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_K823003" style="color:#3ab473">BBa_K823003</a></a></li>
       </ul>
       </ul>
     </ul>
     </ul>
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       <li><a href="https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf" style="color:#f05151"> Transformation</a>:</li>
       <li><a href="https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf" style="color:#f05151"> Transformation</a>:</li>
       <ul>
       <ul>
-
         <li>BBa_316016</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_K316016" style="color:#3ab473">BBa_K316016</a></li>
       </ul>
       </ul>
       <li>Inoculum:</li>
       <li>Inoculum:</li>
       <ul>
       <ul>
-
         <li>BBa_K143055</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_K143055" style="color:#3ab473">BBa_K143055</a></li>
         <li>RBS+qteE</li>
         <li>RBS+qteE</li>
       </ul>
       </ul>
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       <li>Inoculum:</li>
       <li>Inoculum:</li>
       <ul>
       <ul>
-
         <li>BBa_K316016</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_K316016" style="color:#3ab473">BBa_K316016</a></li>
       </ul>
       </ul>
       <li><a href="https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf"style="color:#f05151">Miniprep</a>, <a href="https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf" style="color:#f05151">Restriction analysis</a> and <a href="https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf" style="color:#f05151">Digestion</a>:</li>
       <li><a href="https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf"style="color:#f05151">Miniprep</a>, <a href="https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf" style="color:#f05151">Restriction analysis</a> and <a href="https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf" style="color:#f05151">Digestion</a>:</li>
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       <li>Inoculum</li>
       <li>Inoculum</li>
       <ul>
       <ul>
-
         <li>BBa_K316016</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_K316016" style="color:#3ab473">BBa_K316016</a></li>
       </ul>
       </ul>
       <li>Prepare RBS+lasR and RBS+qteE for sequencing</li>
       <li>Prepare RBS+lasR and RBS+qteE for sequencing</li>
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       <li><a href="https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf"style="color:#f05151">Miniprep</a>:</li>
       <li><a href="https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf"style="color:#f05151">Miniprep</a>:</li>
       <ul>
       <ul>
-
         <li>BBa_K316016</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_K316016" style="color:#3ab473">BBa_K316016</a></li>
       </ul>
       </ul>
     </ul>
     </ul>
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         <li>Inoculum</li>
         <li>Inoculum</li>
       </ul>
       </ul>
-
       <li><a href="https://static.igem.org/mediawiki/2014/7/74/PCR.pdf" style="color:#f05151">PCR</a> BBa_K143055 for RBS removal</li>
+
       <li><a href="https://static.igem.org/mediawiki/2014/7/74/PCR.pdf" style="color:#f05151">PCR</a> <a href="http://parts.igem.org/Part:BBa_K143055" style="color:#3ab473">BBa_K143055</a> for RBS removal</li>
       <li>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 :) </li>
       <li>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 :) </li>
-
       <li>PCR of Plac BBa_K143055 was followed by a <a href="https://static.igem.org/mediawiki/2014/a/af/Agarose_Gel_Electrophoresis_and_DNA_Gel_Purification.pdf"  style="color:#f05151">Gel Purification</a> and <a href="https://static.igem.org/mediawiki/2014/0/03/PCR_Product_Clonig.pdf" style="color:#f05151">Ligation</a> in the PUC19 vector</li>
+
       <li>PCR of Plac <a href="http://parts.igem.org/Part:BBa_K143055" style="color:#3ab473">BBa_K143055</a> was followed by a <a href="https://static.igem.org/mediawiki/2014/a/af/Agarose_Gel_Electrophoresis_and_DNA_Gel_Purification.pdf"  style="color:#f05151">Gel Purification</a> and <a href="https://static.igem.org/mediawiki/2014/0/03/PCR_Product_Clonig.pdf" style="color:#f05151">Ligation</a> in the PUC19 vector</li>
     </ul>
     </ul>
     </td>
     </td>
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       <li><a href="https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf" style="color:#f05151"> Transformation</a>:</li>
       <li><a href="https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf" style="color:#f05151"> Transformation</a>:</li>
       <ul>
       <ul>
-
         <li>BBa_K143055</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_K143055" style="color:#3ab473">BBa_K143055</a></li>
       </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>E. coli DH5-alpha</li>
         <li>E. coli DH5-alpha</li>
         <li><a href="https://static.igem.org/mediawiki/2014/9/90/KI.pdf">KI</a></li>
         <li><a href="https://static.igem.org/mediawiki/2014/9/90/KI.pdf">KI</a></li>
-
         <li>Plac BBa_K143055</li>
+
         <li>Plac <a href="http://parts.igem.org/Part:BBa_K143055" style="color:#3ab473">BBa_K143055</a></li>
       </ul>
       </ul>
       <li><a href="https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf" style="color:#f05151"> Transformation</a></li>
       <li><a href="https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf" style="color:#f05151"> Transformation</a></li>
       <ul>
       <ul>
-
         <li>BBa_P0312</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_P0312" style="color:#3ab473">BBa_P0312</a></li>
       </ul>
       </ul>
       <li>Assembly <a href="https://static.igem.org/mediawiki/2014/f/fe/AV.pdf">AV</a>:</li>
       <li>Assembly <a href="https://static.igem.org/mediawiki/2014/f/fe/AV.pdf">AV</a>:</li>
Line 508: Line 514:
       <li><a href="https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf"style="color:#f05151">Miniprep</a>:</li>
       <li><a href="https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf"style="color:#f05151">Miniprep</a>:</li>
       <ul>
       <ul>
-
         <li>Plac BBa_K143055</li>
+
         <li>Plac <a href="http://parts.igem.org/Part:BBa_K143055" style="color:#3ab473">BBa_K143055</a></li>
       </ul>
       </ul>
       <li>Inoculum</li>
       <li>Inoculum</li>
       <ul>
       <ul>
-
         <li>BBa_P0312</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_P0312" style="color:#3ab473">BBa_P0312</a></li>
       </ul>
       </ul>
       <li>Assembly <a href="https://static.igem.org/mediawiki/2014/f/fe/AV.pdf">AV</a>:</li>
       <li>Assembly <a href="https://static.igem.org/mediawiki/2014/f/fe/AV.pdf">AV</a>:</li>
       <ul>
       <ul>
         <li><a href="https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf"style="color:#f05151">Miniprep</a></li>
         <li><a href="https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf"style="color:#f05151">Miniprep</a></li>
-
         <li>And once again! <a href="https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf" style="color:#f05151">Restriction analysis</a> (this BBa_K316016 is tough)</li>
+
         <li>And once again! <a href="https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf" style="color:#f05151">Restriction analysis</a> (this <a href="http://parts.igem.org/Part:BBa_K316016" style="color:#3ab473">BBa_K316016</a> is tough)</li>
       </ul>
       </ul>
     </ul>
     </ul>
Line 526: Line 532:
       <li><a href="https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf"style="color:#f05151">Miniprep</a> and <a href="https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf" style="color:#f05151">Restriction analysis</a>:</li>
       <li><a href="https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf"style="color:#f05151">Miniprep</a> and <a href="https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf" style="color:#f05151">Restriction analysis</a>:</li>
       <ul>
       <ul>
-
         <li>BBa_P0312</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_P0312" style="color:#3ab473">BBa_P0312</a></li>
       </ul>
       </ul>
     </ul>
     </ul>
Line 536: Line 542:
     <td>11/08
     <td>11/08
     <ul>
     <ul>
-
       <li>Prepare Plac BBa_K143055 for sequencing</li>
+
       <li>Prepare Plac <a href="http://parts.igem.org/Part:BBa_K143055" style="color:#3ab473">BBa_K143055</a> 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>
Line 649: Line 655:
       <li>Inoculum</li>
       <li>Inoculum</li>
       <ul>
       <ul>
-
         <li>BBa_J04450</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_J04450" style="color:#3ab473"> BBa_J04450</a></li>
-
         <li>BBa_K143055</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_K143055" style="color:#3ab473">BBa_K143055</a></li>
       </ul>
       </ul>
-
       <li><a href="https://static.igem.org/mediawiki/2014/7/74/PCR.pdf" style="color:#f05151">PCR</a> BBa_K143055 with higher melting temperature.</li>
+
       <li><a href="https://static.igem.org/mediawiki/2014/7/74/PCR.pdf" style="color:#f05151">PCR</a> <a href="http://parts.igem.org/Part:BBa_K143055" style="color:#3ab473">BBa_K143055</a> with higher melting temperature.</li>
     <ul>
     <ul>
     </td>
     </td>
Line 766: Line 772:
     <td>08/09
     <td>08/09
     <ul>
     <ul>
-
       <li><a href="https://static.igem.org/mediawiki/2014/6/63/QuickChange_PCR.pdf" style="color:#f05151">QuickChange PCR</a> to mutate the cleavage site of the <a href="http://parts.igem.org/Part:BBa_K316037" style="color:#3ab473">BBa_K316037<a/> to a Cathepsin S recognition site</li>
+
       <li><a href="https://static.igem.org/mediawiki/2014/6/63/QuickChange_PCR.pdf" style="color:#f05151">QuickChange PCR</a> to mutate the cleavage site of the <a href="http://parts.igem.org/Part:BBa_K316037" style="color:#3ab473">BBa_K316037</a> to a Cathepsin S recognition site</li>
       <li><a href="https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf" style="color:#f05151"> Transformation</a> of the QuickCHange PCR product</li>
       <li><a href="https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf" style="color:#f05151"> Transformation</a> of the QuickCHange PCR product</li>
     </ul>
     </ul>
Line 774: Line 780:
     <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>
-
       <li><a href="https://static.igem.org/mediawiki/2014/6/63/QuickChange_PCR.pdf" style="color:#f05151">QuickChange PCR</a> to mutate the cleavage site of the <a href="http://parts.igem.org/Part:BBa_K316037" style="color:#3ab473">BBa_K316037<a/> to a Cathepsin S recognition site</li>
+
       <li><a href="https://static.igem.org/mediawiki/2014/6/63/QuickChange_PCR.pdf" style="color:#f05151">QuickChange PCR</a> to mutate the cleavage site of the <a href="http://parts.igem.org/Part:BBa_K316037" style="color:#3ab473">BBa_K316037</a> to a Cathepsin S recognition site</li>
       <li><a href="https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf" style="color:#f05151"> Transformation</a> of the QuickCHange PCR product</li>
       <li><a href="https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf" style="color:#f05151"> Transformation</a> of the QuickCHange PCR product</li>
       <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 807: Line 813:
         <li><a href="https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf" style="color:#f05151">Transformation</a></li>
         <li><a href="https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf" style="color:#f05151">Transformation</a></li>
       </ul>
       </ul>
-
       <li>We had far to many difficulties trying to remove the RBS from the BBa_K143055, so we decided to synthesize it using PCR.</li>
+
       <li>We had far to many difficulties trying to remove the RBS from the <a href="http://parts.igem.org/Part:BBa_K143055" style="color:#3ab473">BBa_K143055</a>, so we decided to synthesize it using PCR.</li>
-
       <li><a href="https://static.igem.org/mediawiki/2014/3/35/PCR_for_synthesis.pdf" style="color:#f05151">PCR for synthesis</a> of the BBa_143015 (BBa_143055 without RBS)</li>
+
       <li><a href="https://static.igem.org/mediawiki/2014/3/35/PCR_for_synthesis.pdf" style="color:#f05151">PCR for synthesis</a> of the <a href="http://parts.igem.org/Part:BBa_K143015" style="color:#3ab473">BBa_K143015</a> (<a href="http://parts.igem.org/Part:BBa_K143055" style="color:#3ab473">BBa_K143055</a> without RBS)</li>
     </ul>
     </ul>
     </td>
     </td>
Line 854: Line 860:
         <li>Inoculum</li>
         <li>Inoculum</li>
       </ul>
       </ul>
-
       <li><a href="https://static.igem.org/mediawiki/2014/a/af/Agarose_Gel_Electrophoresis_and_DNA_Gel_Purification.pdf"  style="color:#f05151">Gel Purification</a> of the BBa_K143015</li>
+
       <li><a href="https://static.igem.org/mediawiki/2014/a/af/Agarose_Gel_Electrophoresis_and_DNA_Gel_Purification.pdf"  style="color:#f05151">Gel Purification</a> of the <a href="http://parts.igem.org/Part:BBa_K143015" style="color:#3ab473">BBa_K143015</a></li>
     </ul>       
     </ul>       
     </td>
     </td>
Line 872: Line 878:
     <td>17/09
     <td>17/09
     <ul>
     <ul>
-
       <li><a href="https://static.igem.org/mediawiki/2014/0/03/PCR_Product_Clonig.pdf" style="color:#f05151">Ligation</a> of the synthesized BBa_K143015 in TOPO vector followed by <a href="https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf" style="color:#f05151"> Transformation</a></li>
+
       <li><a href="https://static.igem.org/mediawiki/2014/0/03/PCR_Product_Clonig.pdf" style="color:#f05151">Ligation</a> of the synthesized <a href="http://parts.igem.org/Part:BBa_K143015" style="color:#3ab473">BBa_K143015</a> in TOPO vector followed by <a href="https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf" style="color:#f05151"> Transformation</a></li>
     </ul>
     </ul>
Line 882: Line 888:
     <td>18/09
     <td>18/09
     <ul>
     <ul>
-
       <li>Replate BBa_K143015. We forgot the X-gal :(</li>
+
       <li>Replate <a href="http://parts.igem.org/Part:BBa_K143015" style="color:#3ab473">BBa_K143015</a>. We forgot the X-gal :(</li>
     </ul>
     </ul>
     <ul>
     <ul>
Line 894: Line 900:
     <td>22/09
     <td>22/09
     <ul>
     <ul>
-
       <li>Inoculum of the BBa_K143015</li>
+
       <li>Inoculum of the <a href="http://parts.igem.org/Part:BBa_K143015" style="color:#3ab473">BBa_K143015</a></li>
       <li>Assembly <a href="https://static.igem.org/mediawiki/2014/6/67/KXIV.pdf">KXIV</a>:</li>
       <li>Assembly <a href="https://static.igem.org/mediawiki/2014/6/67/KXIV.pdf">KXIV</a>:</li>
       <ul>
       <ul>
Line 908: Line 914:
         <li><a href="https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf" style="color:#f05151">Digestion EXSP</a></li>
         <li><a href="https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf" style="color:#f05151">Digestion EXSP</a></li>
       </ul>
       </ul>
-
       <li><a href="https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf">Miniprep</a> and <a href="https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf" style="color:#f05151">Restriction Analysis</a> of the BBa_K143015</li>
+
       <li><a href="https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf">Miniprep</a> and <a href="https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf" style="color:#f05151">Restriction Analysis</a> of the <a href="http://parts.igem.org/Part:BBa_K143015" style="color:#3ab473">BBa_K143015</a></li>
     </ul>
     </ul>
     </td>
     </td>
Line 1,026: Line 1,032:
         <li><a href="https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf" style="color:#f05151">Restriction analysis</a></li>
         <li><a href="https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf" style="color:#f05151">Restriction analysis</a></li>
       </ul>
       </ul>
-
       <li>Assemblies KIX, CIII and KXVI</li>
+
       <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/d/da/KIX.pdf">KIX</a>,
 +
          <a href="https://static.igem.org/mediawiki/2014/5/5b/BIV.pdf">BIV</a> and
 +
          <ahref="https://static.igem.org/mediawiki/2014/6/64/KXVI.pdf">KXVI</a></li>
       <ul>
       <ul>
         <li><a href="https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf" style="color:#f05151">Digestion EXSP</a></li>
         <li><a href="https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf" style="color:#f05151">Digestion EXSP</a></li>
Line 1,032: Line 1,040:
     </ul>
     </ul>
     </td>
     </td>
-
     <td>07/10</td>
+
     <td>07/10  
-
     <td>08/10</td>
+
    <ul>
-
     <td>09/10</td>
+
      <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/d/da/KIX.pdf">KIX</a>,
-
     <td>10/10</td>
+
          <a href="https://static.igem.org/mediawiki/2014/5/5b/BIV.pdf">BIV</a> and
 +
          <a href="https://static.igem.org/mediawiki/2014/6/64/KXVI.pdf">KXVI</a></li>
 +
      <ul>
 +
        <li><a href="https://static.igem.org/mediawiki/2014/a/af/Agarose_Gel_Electrophoresis_and_DNA_Gel_Purification.pdf"  style="color:#f05151">Gel Purification</a></li>
 +
        <li><a href="https://static.igem.org/mediawiki/2014/7/7a/Cohesive-end_assembly_cloning.pdf" style="color:#f05151">Ligation</a></li>
 +
      </ul>
 +
    </ul>
 +
    </td>
 +
     <td>08/10
 +
    <ul>
 +
      <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/d/da/KIX.pdf">KIX</a>,
 +
          <a href="https://static.igem.org/mediawiki/2014/5/5b/BIV.pdf">BIV</a> and
 +
          <a href="https://static.igem.org/mediawiki/2014/6/64/KXVI.pdf">KXVI</a></li>
 +
      <ul>
 +
        <li><a href="https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf" style="color:#f05151"> Transformation</a></li>
 +
      </ul>
 +
    <ul>
 +
    </td>
 +
     <td>09/10
 +
      <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/d/da/KIX.pdf">KIX</a>,
 +
          <a href="https://static.igem.org/mediawiki/2014/5/5b/BIV.pdf">BIV</a> and
 +
          <a href="https://static.igem.org/mediawiki/2014/6/64/KXVI.pdf">KXVI</a></li>
 +
      <ul>
 +
        <li>Inoculum</li>
 +
      </ul>
 +
    </td>
 +
     <td>10/10
 +
    <ul>
 +
      <li>Assemblies <a href="https://static.igem.org/mediawiki/2014/d/da/KIX.pdf">KIX</a>,
 +
          <a href="https://static.igem.org/mediawiki/2014/5/5b/BIV.pdf">BIV</a> and
 +
          <a href="https://static.igem.org/mediawiki/2014/6/64/KXVI.pdf">KXVI</a></li>
 +
      <ul>
 +
        <li><a href="https://static.igem.org/mediawiki/2014/4/42/Restriction_Anaysis_and_Digestion_in_large_scale.pdf" style="color:#f05151">Restriction analysis</a></li>
 +
      </ul>
 +
    </ul>
 +
    </td>
     <td>11/10</td>
     <td>11/10</td>
     <td>12/10</td>
     <td>12/10</td>

Latest revision as of 20:32, 17 October 2014

HeaderNotebookBRASILAP.png

Main Assembly Map

     The main assembly map describes all the constructions that would be done in the project, but some of them have not been performed in time of the iGEM deadline. The succesfully constructed assemblies is indicated by a green check point and the unsuccessfull constructions are indicated by a red "X". Click on these assemblies to access the lab form.

AI AII AIII BIII AIV BII CII DI AVI AV BIV

Assemblies forms

Assembly form template

This is the template designed to help with the laboratory organization during the assemble of biological parts. If you want to use the assembly method we used you can print the form and complete it for each construction. Hope it helps :)

Life Inside the LAB

July

Monday Tuesday Wednesday Thursday Friday Saturday Sunday
30/06
  • PCR lasR (BBa_C0079). The purpose of this PCR was to add the RBS (BBa_K143021), through addition of the sequence in the primer foward, and also to remove the LVA tag. In our primers we only added the restriction sites X and S, so we still need to put it in the pSB1C3 for the Biobrick standard completion. When standardized this part will be called BBa_K1521001.
 01/07 02/07 03/07 04/07 05/07 06/07
07/07 08/07 09/07 10/07
  • Miniprep, Quantification and Restriction Analysis:
    • BBa_B0015 (restriction analysis fail, repeat)
  • PCR the qteE gene for amplification. We also added the RBS (BBa_K143021,) using the foward primer. In our primers we only added the restriction sites X and S, so we still need to put it in the pSB1C3 for the Biobrick standard completion. When standardized this biobrick will be called BBa_K1521000.
  • Ligation of RBS+lasR in PTZ57R/T vector (instaclone kit)
 11/07 12/07 13/07
14/07 15/07 16/07 17/07 18/07 19/07 20/07
21/07 22/07 23/07 24/07 25/07 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 both the construction and the pSB1C3 vector had similar size.
  • Assembly AII, AV and AVI:
 29/07 30/07 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 02/08 03/08
04/08 05//08 06/08 07/08 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 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 17/08
18/08 19/08 20/08
  • RBS+qteE, RBS+lasR and Assemblies BII and BIII:
    • Inoculum
 21/08 22/08 23/08 24/08
25/08 26/08 27/08 28/08 29/08 30/08 31/08



September

Monday Tuesday Wednesday Thursday Friday Saturday Sunday
01/09 02/09 03/09 04/09 05/09 06/09 07/09
08/09 09/09 10/09 11/09 12/09 13/09
  • Assembly KX
    • Inoculum
  • Assemblies DI, KVIII and KXI:
  • Prepare LB medium (solid and liquid)
 14/09
15/09 16/09 17/09
  • Measurement Interlab Study - Samples growth biobrick devices 1, 2 and 3
 18/09
  • Measurement Interlab Study - Sample preparation and characterization by Flow Cytometry and Fluorometry
 19/09 20/09 21/09
22/09 23/09 24/09 25/09
  • Assembly KXIV:
    • Inoculum
 26/09 27/09 28/09
29/09 30/09



October

Monday Tuesday Wednesday Thursday Friday Saturday Sunday
01/10 02/10 03/10 04/10 05/10
06/10 07/10 08/10 09/10
  • Assemblies KIX, BIV and KXVI
    • Inoculum
    		 10/10 11/10 12/10
    13/10 14/10 15/10 16/10 17/10 18/10 19/10
    20/10 21/10 22/10 23/10 24/10 25/10 26/10
    27/10 28/10 29/10 30/10 31/10

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