Team:Freiburg Bioware/NoteBook/Labjournal

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<h2 style="margin-left: 5px;">March</h2>
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<p class="boxes_labjournal_day">Labday 1</p>
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<p class="boxes_labjournal_text">...watching gene sequences...</p>
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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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Vector plasmid; Thymidine kinase ;Cytosindeaminase" />
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<p class="boxes_labjournal_text">...ordering AAV-2 Helper-free System...</p>
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<h2 style="margin-left: 5px;">May</h2>
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<p class="boxes_labjournal_day">Labday 6- 17</p>
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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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<p class="boxes_labjournal_text">...theoretical cloning...</p>
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<p class="boxes_labjournal_day">Labday 18- 45</p>
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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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<h2 style="margin-left: 5px;">July</h2>
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<p class="boxes_labjournal_day">Labday 46- 75</p>
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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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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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<p class="boxes_labjournal_text">...organizing the lab...</p>
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<h2 style="margin-left: 5px;">August I</h2>
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<p class="boxes_labjournal_day">Labday 76- 92</p>
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<p class="boxes_labjournal_text">...cloning parts...</p>
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<h2 style="margin-left: 5px;">August II</h2>
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<p class="boxes_labjournal_day">Labday 93- 106</p>
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<p class="boxes_labjournal_text">...assembled vector plasmid is working...</p>
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<h2 style="margin-left: 5px;">September I</h2>
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<p class="boxes_labjournal_day">Labday 107- 123</p>
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<p class="boxes_labjournal_text">...simply producing BioBricks...</p>
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<h2 style="margin-left: 5px;">September II</h2>
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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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<h2 style="margin-left: 5px;">October I</h2>
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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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<h2 style="margin-left: 5px;">October II</h2>
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<p class="boxes_labjournal_day">Labday 150- 166</p>
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<p class="boxes_labjournal_text">...Capsid modification works...</p>
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<h2 style="margin-left: 5px;">November</h2>
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<p class="boxes_labjournal_day">Labday 166- 170</p>
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<p class="boxes_labjournal_text">...Jamboree!!!...</p>
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<ul>
 
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<li><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/March">March</a></li>
 
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<li><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/April">April</a></li>
 
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<li><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/May">May</a></li>
 
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<li><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/June">June</a></li>
 
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<li><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/July">July</a></li>
 
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<li><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/August">August</a></li>
 
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<li><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/September">September</a></li>
 
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<li><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/October">October</a></li>
 
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<li><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/November">November</a></li>
 
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{{:Team:Freiburg_Bioware/Footer}}

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"