Team:Brasil-SP/Notebook

From 2014.igem.org

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<h1 align=center>Experimental Methodology</h1>
 
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<p>For the assemblies constructuion we addopted the following method:</p>
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<h1 align=center>Main Assembly Map</h1>
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<div align=center><img src="https://static.igem.org/mediawiki/2014/f/fc/Metodologia_Brasil-SP.png" width="900" height="400"/></div>
 
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<br>
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<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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<p>We used the pSB1C3 vector from the Biobrick <a href="http://parts.igem.org/Part:BBa_J04450">BBa_J04450</a>. Using this biobrick we added a red/white selection, where the red colonies are those which do not have the assembly, and the white ones are the correct ones.</p>
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<div align=center><img src="https://static.igem.org/mediawiki/2014/2/25/Red_and_WHite_plate_.jpg" width="700" height="265"/></div>
 
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<p>See our <a href="https://2014.igem.org/Team:Brasil-SP/Notebook/Protocols">Protocol</a>
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  <div class="map_image" style="background-image: url('https://static.igem.org/mediawiki/2014/8/87/Mapa.png');">
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<!--estou colocando os fichamentos dos assemblies aqui, ainda preciso acabar os que faltam mas ja da pra experimentar com a organização destes-->
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<a class="map_link" id="map_link_1" title="" href="https://static.igem.org/mediawiki/2014/b/b7/AI.pdf">AI</a>
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<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_3" title="" href="https://static.igem.org/mediawiki/2014/d/d6/AIII.pdf">AIII</a>
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<a class="map_link" id="map_link_4" title="" href="https://static.igem.org/mediawiki/2014/e/ea/BIII.pdf">BIII</a>
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<a class="map_link" id="map_link_5" title="" href="https://static.igem.org/mediawiki/2014/e/eb/AIV.pdf">AIV</a>
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<a class="map_link" id="map_link_6" title="" href="https://static.igem.org/mediawiki/2014/4/48/BII.pdf">BII</a>
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<a class="map_link" id="map_link_7" title="" href="https://static.igem.org/mediawiki/2014/a/a3/CII.pdf">CII</a>
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<a class="map_link" id="map_link_8" title="" href="https://static.igem.org/mediawiki/2014/7/72/DI.pdf">DI</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>
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<a class="map_link" id="map_link_10" title="" href="https://static.igem.org/mediawiki/2014/f/fe/AV.pdf">AV</a>
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<h1 align=center>Assemblies forms</h1>
<h1 align=center>Assemblies forms</h1>
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<p>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 :)</p>
<p>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 :)</p>
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<a align=center href="https://static.igem.org/mediawiki/2014/e/eb/Assembly_Form.pdf"><img src="https://static.igem.org/mediawiki/2014/3/32/Assembly_form_pg3.png" style="width:700px;height:400px"/></a>
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<a href="https://static.igem.org/mediawiki/2014/e/eb/Assembly_Form.pdf"><img src="https://static.igem.org/mediawiki/2014/3/32/Assembly_form_pg3.png" style="width:700px;height:400px"/></a>
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      <a href="https://static.igem.org/mediawiki/2014/3/38/AII.pdf"><img src="https://static.igem.org/mediawiki/2014/e/e1/AII_image.png" width="100%" height="150px"/></a>
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      <a href="https://static.igem.org/mediawiki/2014/e/eb/AIV.pdf"><img src="https://static.igem.org/mediawiki/2014/4/49/AIV_image.png" width="100%" height="150px"/></a>
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      <a href="https://static.igem.org/mediawiki/2014/d/d6/AVI.pdf"><img src="https://static.igem.org/mediawiki/2014/9/9f/AVI.png"width="100%" height="150px"/></a>
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      <a href="https://static.igem.org/mediawiki/2014/4/48/BII.pdf"><img src="https://static.igem.org/mediawiki/2014/9/9e/BII.png"width="100%" height="150px"/></a>
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      <a href="https://static.igem.org/mediawiki/2014/e/ea/BIII.pdf"><img src="https://static.igem.org/mediawiki/2014/3/36/BIII.png"width="100%" height="150px"/></a>
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      <a href="https://static.igem.org/mediawiki/2014/a/a3/CII.pdf"><img src="https://static.igem.org/mediawiki/2014/3/3a/CII.png"width="100%" height="150px"/></a>
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      <a href="https://static.igem.org/mediawiki/2014/7/72/DI.pdf"><img src="https://static.igem.org/mediawiki/2014/f/f7/DI.png"width="100%" height="150px"/></a>
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<h1>Characterization Assemblies</h1>
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<p> Apart from the main genetic circuit we also assembled others for characterization purposes, such as the validation of the promoters and tunning of our threshold setter concentration, the QteE.</p>
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<h2>Promoter BBa_K823003</h2>
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<p><strong>Question</strong>: Does the constitutive promoter BBa_K823003 work properly?</p>
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<img src="https://static.igem.org/mediawiki/2014/d/d5/KI_w.png" width="300" height="170px"/>
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<p><strong>Results</strong>: After the incubation period of the transformed <em>E. coli</em> a large portion of the colonies were glowing green. So the promoter does work. Moreover, this biobrick works on <em>E. coli</em> despite the fact it was designed for <em>B. subtilis</em>.</p>
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<img src="https://static.igem.org/mediawiki/2014/e/ef/Glowing_E_coli.JPG" width="400px" height="200px"/>
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<h2>Promoter BBa_K143015</h2>
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<p><strong>Question</strong>: How does the transcription caused by this promoter varies with the IPTG induction?</p>
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<img src="https://static.igem.org/mediawiki/2014/7/7a/KXVI_w.png" width="600px" height="350px"/>
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<p><strong>Results</strong>:</p>
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<h2>Tunning of the QteE Threshold</h2>
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<p><strong>Question</strong>:What are the concentration of QteE needed to hamper the LasR induction of the promoter PlasR?</p>
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<p>This is the most difficult task of our project. Tunning the production of QteE so that we establish the correct threshold for the discretization of the Cystatin C level in serum. To attack this challenge we designed 3 circuits so that we could plot a calibration curve. In this circuits we put the transcription of the LasR and QteE under two differnt promoters, the Pveg (BBa_K823003) and PlasR (BBa_K143015).</p>
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<img src="https://static.igem.org/mediawiki/2014/8/86/KXV_w.png" width="600px" height="300px"/>
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<h1>Assembly Forms<h1>
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      <a href="https://static.igem.org/mediawiki/2014/9/90/KI.pdf"><img src="https://static.igem.org/mediawiki/2014/2/23/KI.png"width="100%" height="150px"/></a>
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      <a href="https://static.igem.org/mediawiki/2014/b/b3/KII.pdf"><img src="https://static.igem.org/mediawiki/2014/c/c5/KII.png"width="100%" height="150px"/></a>
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      <a href="https://static.igem.org/mediawiki/2014/7/78/KIII.pdf"><img src="https://static.igem.org/mediawiki/2014/e/ed/KIII.png"width="100%" height="150px"/></a>
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      <a href="https://static.igem.org/mediawiki/2014/9/95/KIV.pdf"><img src="https://static.igem.org/mediawiki/2014/e/ee/KIV.png"width="100%" height="150px"/></a>
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      <a href="https://static.igem.org/mediawiki/2014/3/3d/KV.pdf"><img src="https://static.igem.org/mediawiki/2014/a/ac/KV.png"width="100%" height="150px"/></a>
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      <a href="https://static.igem.org/mediawiki/2014/8/83/KVI.pdf"><img src="https://static.igem.org/mediawiki/2014/3/37/KVI.png"width="100%" height="150px"/></a>
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      <a href="https://static.igem.org/mediawiki/2014/9/95/KX.pdf"><img src="https://static.igem.org/mediawiki/2014/d/d5/KX.png"width="100%" height="150px"/></a>
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      <a href="https://static.igem.org/mediawiki/2014/9/9c/KXI.pdf"><img src="https://static.igem.org/mediawiki/2014/b/b8/KXI.png"width="100%" height="150px"/></a>
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</table>-->
 
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      <li><a href="https://static.igem.org/mediawiki/2014/b/b7/AI.pdf">AI</a></li>
 
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<h1 align=center><strong>Life Inside the LAB</strong></h1>
<h1 align=center><strong>Life Inside the LAB</strong></h1>
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<thead><h2>July</h2>
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<thead><h2 align=center>July</h2>
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     <td>30/06
     <td>30/06
     <ul>
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       <li><a href="https://static.igem.org/mediawiki/2014/7/74/PCR.pdf" style="color:#f05151">PCR</a> 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.</li>
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       <li><a href="https://static.igem.org/mediawiki/2014/7/74/PCR.pdf" style="color:#f05151">PCR</a> lasR (<a href="http://parts.igem.org/Part:BBa_C0079" style="color:#3ab473">BBa_C0079</a>). The purpose of this PCR was to add the RBS (<a href="http://parts.igem.org/Part:BBa_K143021" style="color:#3ab473">BBa_K143021</a>), 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 <a href="http://parts.igem.org/Part:BBa_K1521001" style="color:#3ab473">BBa_K1521001</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> of the Biobricks:</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 Biobricks:</li>
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         <li><a href="http://parts.igem.org/Part:BBa_K143012" style="color:#3ab473">BBa_K143012</a></li>
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         <li><a href="http://parts.igem.org/Part:BBa_E0840" style="color:#3ab473">BBa_E0840</a></li>
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       <li>Inoculum of the Biobricks sent by iGEM HQ:</li>
       <li>Inoculum of the Biobricks sent by iGEM HQ:</li>
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         <li><a href="http://parts.igem.org/Part:BBa_K316037" style="color:#3ab473">BBa_K316037<a/></li>
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         <li>BBa_K316015</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_K316015" style="color:#3ab473">BBa_K316015<a/></li>
       </ul>
       </ul>
     </ul>
     </ul>
Line 307: Line 155:
       <li><a href="https://static.igem.org/mediawiki/2014/a/a5/Transformation_in_Escherichia_coli_DH5.pdf" style="color:#f05151"> Transformation</a> of the Biobricks sent by the Imperial College Team:</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 Biobricks sent by the Imperial College Team:</li>
       <ul>
       <ul>
-
         <li>BBa_K316016</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_K316016" style="color:#3ab473">BBa_K316016<a/></li>
-
         <li>BBa_K143031</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_K143031" style="color:#3ab473">BBa_K143031<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>, Quantification of the plasmidial DNA samples 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>, Quantification of the plasmidial DNA samples 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_143012</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_K316037" style="color:#3ab473">BBa_K316037<a/></li>
-
         <li>BBa_K081001</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_K316018" style="color:#3ab473">BBa_K316018<a/></li>
-
         <li>BBa_E0840</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_K316015" style="color:#3ab473">BBa_K316015<a/></li>
       </ul>
       </ul>
     </ul>
     </ul>
Line 324: Line 172:
       <li>Inoculum:</li>
       <li>Inoculum:</li>
       <ul>
       <ul>
-
         <li>BBa_143012</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_K143012" style="color:#3ab473">BBa_K143012</a></li>
-
         <li>BBa_K081001</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_K081001" style="color:#3ab473">BBa_K081001</a></li>
-
         <li>BBa_E0840</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_E0840" style="color:#3ab473">BBa_E0840</a></li>
       </ul>
       </ul>
     </ul>
     </ul>
Line 336: Line 184:
       <li><a href="https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf"style="color:#f05151">Miniprep</a>, Quantification 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>, Quantification 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_143012</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_K143012" style="color:#3ab473">BBa_K143012</a></li>
-
         <li>BBa_K081001</li>
+
         <li><a href="http://parts.igem.org/Part:BBa_K081001" style="color:#3ab473">BBa_K081001</a></li>
-
         <li>BBa_E0840</li>
+
         <li<a href="http://parts.igem.org/Part:BBa_E0840" style="color:#3ab473">BBa_E0840</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_B0015</li>
+
         <li><a href"http://parts.igem.org/Part:BBa_B0015" style="color:#3ab473">BBa_B0015</a></li>
       </ul>
       </ul>
     </ul>
     </ul>
Line 351: Line 199:
     <li>Inoculum:</li>
     <li>Inoculum:</li>
       <ul>
       <ul>
-
         <li>BBa_B0015</li>
+
         <li><a href"http://parts.igem.org/Part:BBa_B0015" style="color:#3ab473">BBa_B0015</a></li>
       </ul>
       </ul>
     </ul>
     </ul>
Line 359: Line 207:
       <li><a href="https://static.igem.org/mediawiki/2014/6/6f/Purelink_quick_plasmid_qrc.pdf"style="color:#f05151">Miniprep</a>, Quantification 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>, Quantification 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_B0015 (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 (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.</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>
Line 370: Line 218:
       <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>
Line 401: Line 249:
       <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>
       <ul>
       <ul>
-
         <li>BBa_B0015</li>
+
         <li><a href"http://parts.igem.org/Part:BBa_B0015" style="color:#3ab473">BBa_B0015</a></li>
       </ul>
       </ul>
     </ul>
     </ul>
Line 413: Line 261:
     <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>
     <li>Electrophoresis analysis:</li>
     <li>Electrophoresis analysis:</li>
       <ul>
       <ul>
-
       <li>BBa_B0015</li>
+
       <li><a href"http://parts.igem.org/Part:BBa_B0015" style="color:#3ab473">BBa_B0015</a></li>
       </ul>
       </ul>
     </ul>
     </ul>
Line 426: Line 274:
       <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>BBa_B0015</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/2014/4/42/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>
Line 449: Line 297:
       <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>BBa_B0015</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>
Line 470: Line 318:
       <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>
Line 487: Line 335:
       <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>
Line 510: Line 358:
       <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>
Line 523: Line 371:
       <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>
Line 584: Line 432:
         <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>
Line 602: Line 450:
</tbody>
</tbody>
-
<thead><h2>August</h2>
+
<thead><h2 align=center>August</h2>
</thead>
</thead>
Line 626: Line 474:
       <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>,
Line 648: Line 496:
         <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 666: 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 684: 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 694: 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 807: 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 847: Line 695:
<br>
<br>
-
<thead><h2>September</h2>
+
<thead><h2 align=center>September</h2>
</thead>
</thead>
Line 924: 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 BBa_316037 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 932: 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 BBa_316037 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 945: Line 793:
     <td>11/09
     <td>11/09
     <ul>
     <ul>
-
       <li>Prepare inoculum of the mutated BBa_K316037</li>
+
       <li>Prepare inoculum of the mutated <a href="http://parts.igem.org/Part:BBa_K316037" style="color:#3ab473">BBa_K316037<a/></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>,  
           <a href="https://static.igem.org/mediawiki/2014/c/ce/KVIII.pdf">KVIII</a>,  
           <a href="https://static.igem.org/mediawiki/2014/c/ce/KVIII.pdf">KVIII</a>,  
Line 965: 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 1,012: 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 1,030: 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 1,040: 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 1,052: 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 1,066: 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 1,141: Line 989:
-
<thead><h2>October</h2>
+
<thead><h2 align=center>October</h2>
</thead>
</thead>
Line 1,184: 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 1,190: 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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