Team:Freiburg Bioware/NoteBook/Labjournal

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<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/March">
<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/March">
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<h2>March</h2>
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<h2 style="margin-left: 5px;">March</h2>
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<p class="boxes_labjournal_text">Labday 1</p>
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<p class="boxes_labjournal_day">Labday 1</p>
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<img src="https://static.igem.org/mediawiki/2010/b/b4/Freiburg_10_labjournal_march_v1.JPG" class="image_labjournal" title="Our project starts with first sequence analysis."/>
<p class="boxes_labjournal_text">...watching gene sequences...</p>
<p class="boxes_labjournal_text">...watching gene sequences...</p>
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<img src="https://static.igem.org/mediawiki/2010/3/35/Freiburg_10_BioBrick_icon.png" class="image_labjournal" />
 
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<h2 style="margin-left: 5px;">April</h2>
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<p class="boxes_labjournal_day">Labday 2- 5</p>
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<img src="https://static.igem.org/mediawiki/2010/f/fc/Freiburg_10_labjournal_april_v1.JPG" class="image_labjournal" title="Scanning for iGEM restriction sites:
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<div class="lab_journal_box">
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Vector plasmid; Thymidine kinase ;Cytosindeaminase" />
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<div class="lab_journal_text">
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<p class="boxes_labjournal_text">...ordering AAV-2 Helper-free System...</p>
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<br><h2>Get to know our virus</h2>
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</a>
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<!---Insert Text in here---> Our project starts with first sequence analysis.
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<div class="lab_journal_img"><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/March" ><img src="https://static.igem.org/mediawiki/2010/2/21/Freiburg10_Labjournal-March.png" /></a></div></div>
 
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<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/May">
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<br><h2>Checking iGEM compatibility </h2>
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<h2 style="margin-left: 5px;">May</h2>
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<!---Insert Text in here--->Scanning for iGEM restriction sites:<br/>
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<p class="boxes_labjournal_day">Labday 6- 17</p>
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• Vector plasmid <br/>
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<img src="https://static.igem.org/mediawiki/2010/0/0f/Freiburg_10_labjournal_may_v1.JPG" class="image_labjournal" title="We took a closer look to the theoretical DNA- and amino acid sequence of the ITRs, questioned the role of the β-globin intron, had a closer look to the CMV promoter and performed our first cloning attempt." />
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• Thymidine kinase<br/>
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<p class="boxes_labjournal_text">...theoretical cloning...</p>
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• Cytosindeaminase<br/>
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</a>
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<div class="lab_journal_img"><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/April" ><img src="https://static.igem.org/mediawiki/2010/6/6b/Freiburg10_Labjournal-April.png" /></a></div></div>
 
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<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/June">
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<br><h2>First practical steps</h2>
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<h2 style="margin-left: 5px;">June</h2>
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<!---Insert Text in here--->We took a closer look to the theoretical DNA- and amino acid sequence of the ITRs, questioned the role of the β-globin intron, had a closer look to the CMV promoter and performed our first cloning attempt.  
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<p class="boxes_labjournal_day">Labday 18- 45</p>
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<img src="https://static.igem.org/mediawiki/2010/3/37/Freiburg_10_labjournal_june_v1.jpg" class="image_labjournal" title="
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We conducted our first site-directed mutagenesis. First cell culture steps: Splitting and seeding of AAV 293 and HT1080 cells; Calcium phosphate transfection; Transduction with virus particles containing YFP encoding sequence; Microscopy: Fluorescent transduced HT1080 cells. We planned to determine infectious virus titer via quantitative real-time PCR. For this purpose primers were designed, transduced HT1080 cells were harvested and prepared. Theoretical studies of the AAV structure: Impressions" />
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<p class="boxes_labjournal_text">...first transduced cells...</p>
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</a>
</div>
</div>
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<div class="lab_journal_img"><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/May" ><img src="https://static.igem.org/mediawiki/2010/6/61/Freiburg10_Labjournal-May.png" /></a></div></div>
 
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<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/July">
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<br><h2>First fluorescing HT1080 cells</h2>
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<h2 style="margin-left: 5px;">July</h2>
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<!---Insert Text in here--->We conducted our first site-directed mutagenesis. <br/>
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<p class="boxes_labjournal_day">Labday 46- 75</p>
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<br/>
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<img src="https://static.igem.org/mediawiki/2010/8/8d/Freiburg_10_labjournal_july_v1.JPG" class="image_labjournal" title="We developed a plan for modifying the surface exposed loops of the virus capsid.
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First cell culture steps:<br/>
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For this purpose not only 5 iGEM restriction sites, but also the so called ViralBrick restriction sites located in the Rep/Cap sequence needed to be deleted: BamHI, SalI. BioBrick production of the inverted terminal repeats (ITRs) turned out to be difficult. Therefore the alternative “ITR fancy method” was developed. We successfully produced AAV2 left and right ITR fused to the RFC10 prefix. Further on we received first quantitative transduction results via flow cytometry. mGMK_TK30 was successfully cloned into the vector plasmid." />
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• Splitting and seeding of AAV 293 and HT1080 cells<br/>
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<p class="boxes_labjournal_text">...organizing the lab...</p>
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• Calcium phosphate transfection<br/>
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</a>
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•      Transduction with virus particles containing YFP encoding sequence<br/>
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-
• Microscopy: Fluorescent transduced HT1080 cells<br/>
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-
<br/>
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We planned to determine infectious virus titer via quantitative real-time PCR. For this purpose primers were designed, transduced HT1080 cells were harvested and prepared.<br/>
+
-
<br/>
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-
Theoretical studies of the AAV structure: Impressions.
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</div>
</div>
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<div class="lab_journal_img"><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/June" ><img src="https://static.igem.org/mediawiki/2010/4/48/Freiburg10_Labjournal-June.png" /></a></div></div>
 
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<br><h2>Modifying Rep/Cap & Modularizing Vector plasmid</h2>
 
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<!---Insert Text in here--->We developed a plan for modifying the surface exposed loops of the virus capsid.
 
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For this purpose not only 5 iGEM restriction sites, but also the so called ViralBrick restriction sites located in the Rep/Cap sequence needed to be deleted: BamHI, SalI <br/>
 
-
<br/>
 
-
BioBrick production of the inverted terminal repeats (ITRs) turned out to be difficult. Therefore the alternative “ITR fancy method” was developed. We successfully produced AAV2 left and right ITR fused to the RFC10 prefix.<br/>
+
<div class="box_labjournal_195">
-
<br/>
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<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/August">
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Further on we received first quantitative transduction results via flow cytometry.<br/>
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<h2 style="margin-left: 5px;">August I</h2>
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<br/>
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<p class="boxes_labjournal_day">Labday 76- 92</p>
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mGMK_TK30 was successfully cloned into the vector plasmid.
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<img src="https://static.igem.org/mediawiki/2010/8/8b/Freiburg_10_labjournal_august1_v1.JPG" class="image_labjournal" />
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<p class="boxes_labjournal_text">...cloning parts...</p>
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</a>
</div>
</div>
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<div class="lab_journal_img"><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/July" ><img src="https://static.igem.org/mediawiki/2010/d/dc/Freiburg10_Labjournal-July.png" /></a></div></div>
 
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<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/August2">
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<br><h2>Producing more BioBricks</h2>
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<h2 style="margin-left: 5px;">August II</h2>
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<!---Insert Text in here--->  
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<p class="boxes_labjournal_day">Labday 93- 106</p>
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<img src="https://static.igem.org/mediawiki/2010/e/ec/Freiburg_10_labjournal_august2_v1.png" class="image_labjournal" />
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<p class="boxes_labjournal_text">...assembled vector plasmid is working...</p>
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</a>
</div>
</div>
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<div class="lab_journal_img"><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/August" ><img src="https://static.igem.org/mediawiki/2010/0/06/Freiburg10_Labjournal-AugustI.png" /></a></div></div>
 
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<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/September">
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<br><h2>Headline</h2>
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<h2 style="margin-left: 5px;">September I</h2>
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<!---Insert Text in here--->  
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<p class="boxes_labjournal_day">Labday 107- 123</p>
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<img src="https://static.igem.org/mediawiki/2010/2/25/Freiburg_10_labjournal_september1_v1.JPG" class="image_labjournal" />
 +
<p class="boxes_labjournal_text">...simply producing BioBricks...</p>
 +
</a>
</div>
</div>
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<div class="lab_journal_img"><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/August2" ><img src="https://static.igem.org/mediawiki/2010/b/b1/Freiburg10_Labjournal-AugustII.png" /></a></div></div>
 
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<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/September2">
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<br><h2>Headline</h2>
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<h2 style="margin-left: 5px;">September II</h2>
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<!---Insert Text in here--->  
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<p class="boxes_labjournal_day">Labday 124- 135</p>
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<img src="https://static.igem.org/mediawiki/2010/1/18/Freiburg_10_labjournal_october2_v1.JPG" class="image_labjournal" />
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<p class="boxes_labjournal_text">...21x mutated RepCap is working...</p>
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</a>
</div>
</div>
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<div class="lab_journal_img"><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/September" ><img src="https://static.igem.org/mediawiki/2010/1/1e/Freiburg10_Labjournal-SeptemberI.png" /></a></div></div>
 
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<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/October">
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<br><h2>Headline</h2>
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<h2 style="margin-left: 5px;">October I</h2>
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<!---Insert Text in here--->  
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<p class="boxes_labjournal_day">Labday 136- 149</p>
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<img src="https://static.igem.org/mediawiki/2010/e/e0/Freiburg_10_labjournal_october_v1.JPG" class="image_labjournal" />
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<p class="boxes_labjournal_text">...testing BioBricks by virus production...</p>
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</a>
</div>
</div>
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<div class="lab_journal_img"><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/September2" ><img src="https://static.igem.org/mediawiki/2010/2/2e/Freiburg10_Labjournal-SeptemberII.png" /></a></div></div>
 
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<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/October2">
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<br><h2>Headline</h2>
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<h2 style="margin-left: 5px;">October II</h2>
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<!---Insert Text in here--->  
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<p class="boxes_labjournal_day">Labday 150- 166</p>
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<img src="https://static.igem.org/mediawiki/2010/2/22/Freiburg10_Oktober.png" class="image_labjournal" />
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<p class="boxes_labjournal_text">...Capsid modification works...</p>
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</a>
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<div class="lab_journal_img"><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/October" ><img src="https://static.igem.org/mediawiki/2010/6/65/Freiburg10_Labjournal-OctoberI.png" /></a></div></div>
 
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<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/November">
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<br><h2>Headline</h2>
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<h2 style="margin-left: 5px;">November</h2>
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<!---Insert Text in here--->  
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<p class="boxes_labjournal_day">Labday 166- 170</p>
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<img src="https://static.igem.org/mediawiki/2010/9/93/Freiburg10_November.png" class="image_labjournal" />
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<p class="boxes_labjournal_text">...Jamboree!!!...</p>
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<div class="lab_journal_img"><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/October2" ><img src="https://static.igem.org/mediawiki/2010/4/48/Freiburg10_Labjournal-June.png" /></a></div></div>
 
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https://2010.igem.org/Team:Freiburg_Bioware/NoteBook => Back to Notebook overview]
 
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<li><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/March">March (labday 1)</a></li>
 
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<li><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/April">April (labday 2 - 5)</a></li>
 
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<li><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/May">May (labday 6 - 17)</a></li>
 
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<li><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/June">June (labday 18 - 45)</a></li>
 
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<li><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/July">July (labday 46 - 75)</a></li>
 
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<li><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/August">August part 1 (labday 76 - 92)</a></li>
 
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<li><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/August2">August part 2 (labday 93 - 106)</a></li>
 
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<li><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/September">September part 1 (labday 107 - 123)</a></li>
 
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<li><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/September2">September part 2 (labday 124 - 135)</a></li>
 
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<li><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/October">October part 1 (labday 136 - 149 )</a></li>
 
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<li><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/October2">October part 2 (labday 150 - 166 )</a></li>
 
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<li><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/November">November  (labday 167 - 170 )</a></li>
 
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<li><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/Cellculture">Cellculture</a></li>
 
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{{:Team:Freiburg_Bioware/Footer}}
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Latest revision as of 03:17, 28 October 2010

March

Labday 1

...watching gene sequences...

April

Labday 2- 5

...ordering AAV-2 Helper-free System...

May

Labday 6- 17

...theoretical cloning...

June

Labday 18- 45

...first transduced cells...

July

Labday 46- 75

...organizing the lab...

August I

Labday 76- 92

...cloning parts...

August II

Labday 93- 106

...assembled vector plasmid is working...

September I

Labday 107- 123

...simply producing BioBricks...

September II

Labday 124- 135

...21x mutated RepCap is working...

October I

Labday 136- 149

...testing BioBricks by virus production...

October II

Labday 150- 166

...Capsid modification works...

November

Labday 166- 170

...Jamboree!!!...

Retrieved from "http://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal"