Team:Freiburg Bioware/NoteBook/Labjournal

From 2010.igem.org

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<div class="box_labjournal_195">
<div class="box_labjournal_195">
<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>Labday 1<br />...watching gene sequences...</p>
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<p class="boxes_labjournal_day">Labday 1</p>
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<img class="image_labjournal" src="https://static.igem.org/mediawiki/2010/3/35/Freiburg_10_BioBrick_icon.png" />
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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>
</a>
</a>
</div>
</div>
<div class="box_labjournal_195">
<div class="box_labjournal_195">
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<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/March">
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<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/April">
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<h2>March</h2>
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<h2 style="margin-left: 5px;">April</h2>
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<p>Labday 1<br />...watching gene sequences...</p>
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<p class="boxes_labjournal_day">Labday 2- 5</p>
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<img class="image_labjournal" src="https://static.igem.org/mediawiki/2010/3/35/Freiburg_10_BioBrick_icon.png" />
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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:
 +
Vector plasmid; Thymidine kinase ;Cytosindeaminase" />
 +
<p class="boxes_labjournal_text">...ordering AAV-2 Helper-free System...</p>
</a>
</a>
</div>
</div>
<div class="box_labjournal_195">
<div class="box_labjournal_195">
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<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/March">
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<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/May">
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<h2>March</h2>
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<h2 style="margin-left: 5px;">May</h2>
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<p>Labday 1<br />...watching gene sequences...</p>
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<p class="boxes_labjournal_day">Labday 6- 17</p>
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<img class="image_labjournal" src="https://static.igem.org/mediawiki/2010/3/35/Freiburg_10_BioBrick_icon.png" />
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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." />
 +
<p class="boxes_labjournal_text">...theoretical cloning...</p>
</a>
</a>
</div>
</div>
<div class="box_labjournal_195">
<div class="box_labjournal_195">
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<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/March">
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<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/June">
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<h2>March</h2>
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<h2 style="margin-left: 5px;">June</h2>
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<p>Labday 1<br />...watching gene sequences...</p>
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<p class="boxes_labjournal_day">Labday 18- 45</p>
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<img class="image_labjournal" src="https://static.igem.org/mediawiki/2010/3/35/Freiburg_10_BioBrick_icon.png" />
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<img src="https://static.igem.org/mediawiki/2010/3/37/Freiburg_10_labjournal_june_v1.jpg" class="image_labjournal" title="
 +
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" />
 +
<p class="boxes_labjournal_text">...first transduced cells...</p>
</a>
</a>
</div>
</div>
<div class="box_labjournal_195">
<div class="box_labjournal_195">
-
<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/March">
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<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/July">
-
<h2>March</h2>
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<h2 style="margin-left: 5px;">July</h2>
-
<p>Labday 1<br />...watching gene sequences...</p>
+
<p class="boxes_labjournal_day">Labday 46- 75</p>
-
<img class="image_labjournal" src="https://static.igem.org/mediawiki/2010/3/35/Freiburg_10_BioBrick_icon.png" />
+
<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.
 +
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." />
 +
<p class="boxes_labjournal_text">...organizing the lab...</p>
</a>
</a>
</div>
</div>
<div class="box_labjournal_195">
<div class="box_labjournal_195">
-
<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/March">
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<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/August">
-
<h2>March</h2>
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<h2 style="margin-left: 5px;">August I</h2>
-
<p>Labday 1<br />...watching gene sequences...</p>
+
<p class="boxes_labjournal_day">Labday 76- 92</p>
-
<img class="image_labjournal" src="https://static.igem.org/mediawiki/2010/3/35/Freiburg_10_BioBrick_icon.png" />
+
<img src="https://static.igem.org/mediawiki/2010/8/8b/Freiburg_10_labjournal_august1_v1.JPG" class="image_labjournal" />
 +
<p class="boxes_labjournal_text">...cloning parts...</p>
</a>
</a>
</div>
</div>
<div class="box_labjournal_195">
<div class="box_labjournal_195">
-
<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/March">
+
<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/August2">
-
<h2>March</h2>
+
<h2 style="margin-left: 5px;">August II</h2>
-
<p>Labday 1<br />...watching gene sequences...</p>
+
<p class="boxes_labjournal_day">Labday 93- 106</p>
-
<img class="image_labjournal" src="https://static.igem.org/mediawiki/2010/3/35/Freiburg_10_BioBrick_icon.png" />
+
<img src="https://static.igem.org/mediawiki/2010/e/ec/Freiburg_10_labjournal_august2_v1.png" class="image_labjournal" />
 +
<p class="boxes_labjournal_text">...assembled vector plasmid is working...</p>
</a>
</a>
</div>
</div>
<div class="box_labjournal_195">
<div class="box_labjournal_195">
-
<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/March">
+
<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/September">
-
<h2>March</h2>
+
<h2 style="margin-left: 5px;">September I</h2>
-
<p>Labday 1<br />...watching gene sequences...</p>
+
<p class="boxes_labjournal_day">Labday 107- 123</p>
-
<img class="image_labjournal" src="https://static.igem.org/mediawiki/2010/3/35/Freiburg_10_BioBrick_icon.png" />
+
<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>
</a>
</div>
</div>
<div class="box_labjournal_195">
<div class="box_labjournal_195">
-
<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/March">
+
<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/September2">
-
<h2>March</h2>
+
<h2 style="margin-left: 5px;">September II</h2>
-
<p>Labday 1<br />...watching gene sequences...</p>
+
<p class="boxes_labjournal_day">Labday 124- 135</p>
-
<img class="image_labjournal" src="https://static.igem.org/mediawiki/2010/3/35/Freiburg_10_BioBrick_icon.png" />
+
<img src="https://static.igem.org/mediawiki/2010/1/18/Freiburg_10_labjournal_october2_v1.JPG" class="image_labjournal" />
 +
<p class="boxes_labjournal_text">...21x mutated RepCap is working...</p>
</a>
</a>
</div>
</div>
-
<!-- Boxes on the right starts -->
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<div class="box_labjournal_195">
-
 
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<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/October">
-
<div class="lab_journal_box">
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<h2 style="margin-left: 5px;">October I</h2>
-
<div class="lab_journal_text">
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<p class="boxes_labjournal_day">Labday 136- 149</p>
-
<br><h2>Get to know our virus</h2>
+
<img src="https://static.igem.org/mediawiki/2010/e/e0/Freiburg_10_labjournal_october_v1.JPG" class="image_labjournal" />
-
<!---Insert Text in here---> Our project starts with first sequence analysis.
+
<p class="boxes_labjournal_text">...testing BioBricks by virus production...</p>
 +
</a>
</div>
</div>
-
<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>
 
-
<div class="lab_journal_box">
+
<div class="box_labjournal_195">
-
<div class="lab_journal_text">
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<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/October2">
-
<br><h2>Checking iGEM compatibility </h2>
+
<h2 style="margin-left: 5px;">October II</h2>
-
<!---Insert Text in here--->Scanning for iGEM restriction sites:<br/>
+
<p class="boxes_labjournal_day">Labday 150- 166</p>
-
• Vector plasmid <br/>
+
<img src="https://static.igem.org/mediawiki/2010/2/22/Freiburg10_Oktober.png" class="image_labjournal" />
-
• Thymidine kinase<br/>
+
<p class="boxes_labjournal_text">...Capsid modification works...</p>
-
• Cytosindeaminase<br/>
+
</a>
-
+
</div>
</div>
-
<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>
 
-
<div class="lab_journal_box">
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<div class="box_labjournal_195">
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<div class="lab_journal_text">
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<a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/November">
-
<br><h2>First practical steps</h2>
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<h2 style="margin-left: 5px;">November</h2>
-
<!---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.
+
<p class="boxes_labjournal_day">Labday 166- 170</p>
 +
<img src="https://static.igem.org/mediawiki/2010/9/93/Freiburg10_November.png" class="image_labjournal" />
 +
<p class="boxes_labjournal_text">...Jamboree!!!...</p>
 +
</a>
</div>
</div>
-
<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>
 
-
<div class="lab_journal_box">
 
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<div class="lab_journal_text">
 
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<br><h2>First fluorescing HT1080 cells</h2>
 
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<!---Insert Text in here--->We conducted our first site-directed mutagenesis. <br/>
 
-
<br/>
 
-
First cell culture steps:<br/>
 
-
• Splitting and seeding of AAV 293 and HT1080 cells<br/>
 
-
• Calcium phosphate transfection<br/>
 
-
•      Transduction with virus particles containing YFP encoding sequence<br/>
 
-
• Microscopy: Fluorescent transduced HT1080 cells<br/>
 
-
<br/>
 
-
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/>
 
-
Theoretical studies of the AAV structure: Impressions.
 
-
 
</div>
</div>
-
<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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<div class="lab_journal_box">
 
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<div class="lab_journal_text">
 
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<br><h2>Modifying Rep/Cap & Modularizing Vector plasmid</h2>
 
-
<!---Insert Text in here--->We developed a plan for modifying the surface exposed loops of the virus capsid.
 
-
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/>
 
-
<br/>
 
-
Further on we received first quantitative transduction results via flow cytometry.<br/>
 
-
<br/>
 
-
mGMK_TK30 was successfully cloned into the vector plasmid.
 
-
 
-
</div>
 
-
<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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<br><h2>Producing more BioBricks</h2>
 
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<!---Insert Text in here--->
 
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</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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<br><h2>Headline</h2>
 
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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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<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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<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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<div class="lab_journal_img"><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/November" ><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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<ul>
 
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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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Latest revision as of 03:17, 28 October 2010