Team:Goettingen/week7-2

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Language: <img height="20", src="http://www.patrickreinke.de/igem/eng.jpg"> English, <img height="20", src="http://www.patrickreinke.de/igem/deu.jpg"> <a href="https://2012.igem.org/Team:Goettingen/main_deu">Deutsch</a> <br>
Language: <img height="20", src="http://www.patrickreinke.de/igem/eng.jpg"> English, <img height="20", src="http://www.patrickreinke.de/igem/deu.jpg"> <a href="https://2012.igem.org/Team:Goettingen/main_deu">Deutsch</a> <br>
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<table id="toc" class="toc"><tbody><tr><td><div id="toctitle"><h2>Contents</h2> <span class="toctoggle">[<a href="javascript:toggleToc()" class="internal" id="togglelink">hide</a>]</span></div>
 
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<ul>
 
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<li class="toclevel-1"><a href="#week7"><span class="tocnumber">1</span> <span class="toctext">7th week (11th to 17th June)</span></a></li>
 
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<li class="toclevel-1"> <div style="text-indent:10px;"><a href="#week7#1"><span class="tocnumber"></span> <span class="toctext">#1 Selection / Swimming</span></a></li>
 
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<li class="toclevel-1"> <div style="text-indent:10px;"><a href="#week7#2"><span class="tocnumber"></span> <span class="toctext">#2 Speed Improvement</span></a></li>
 
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<li class="toclevel-1"> <div style="text-indent:10px;"><a href="#week7#3"><span class="tocnumber"></span> <span class="toctext">#3 Chemoreceptor Library</span></a></li>
 
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<h2><b><a name="week7#1">#1 Selection / Swimming</a></b></h2>
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<h2><b><a name="week6#2">#2 Speed Improvement - 6th week</a></b></h2>
<a href="https://2012.igem.org/Team:Goettingen/Notebook">Back to overview</a><br>
<a href="https://2012.igem.org/Team:Goettingen/Notebook">Back to overview</a><br>
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Under process..
 
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<h2><b></b></h2><br>
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<h2><b>V06_04 </b></h2><br>
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<b></b><br>
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<b>Chemical retransformartion of the <i>flhDC</i>-promoter constructs into <i>E. coli</i> (DH10B)</i></b><br>
<ul>
<ul>
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<li>Experiment: <br> </li>
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<li>Experiment: <br>  
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<li>Description: <br> </li>
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For the chemical retransformation the standard protocol for transformation was followed. The following constructs were transformed into
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<li>Observations & Results: <br> </li>
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E. coli: <br>
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</ul>
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20E - #2 <br>
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20E - #3 <br>
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<b><i/></b><br>
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18M - #2 <br>
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18M - #3 <br>
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<li>Experiment: </li>
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18C - #1 <br>
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<li>Description: <br> </li>
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18C - #2 <br></li>  
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<li>Observations & Results: <br> </li>
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<li>Observations & Results: <br>
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The retransformation was successful since all plates showed numeours colonoes except the negative control.
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<br> </li>
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<h2><b><a name="week7#2">#2 Speed Improvement</a></b></h2>
 
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<a href="https://2012.igem.org/Team:Goettingen/Notebook">Back to overview</a><br>
 
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Under process...
 
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<h2><b>V06_05 </b></h2><br>
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<b>V06_05_1 Preparative double digestion of <i>flhDC</i> and plasmids containing further promoter</b><br>
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<li>Experiment:  <br> </li>
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<li>Experiment:  <br>
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<li>Description: <br> </li>
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In order to clone <i>flhDC</i> behind further promoters, the purified PCR product was digested with XbaI and PstI, whereas the BioBrick plasmids were cut with SpeI and PstI. The digestion was performed as described in the protocol. This time the follwing promoters were used: <br>
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<li>Observations & Results: <br> </li>
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J23113 - 20G - x21 <br>
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J23109  - 2G - x106 <br>
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J23114 - 20I - x256 <br>
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J23106 - 18O - x1185 <br>
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J23104 - 18K - x 1831 <br>
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<b><i/></b><br>
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<b>V06_05_2  Insertion of <i>flhDC</i> into the BioBrick plamids</b><br>
<ul>
<ul>
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<li>Experiment: </li>
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<li>Experiment: <br>
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<li>Description: <br> </li>
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The ligation of the digested flhDC with the BioBrick plasmids was conducted as described in the protocol. <br> </li>
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<li>Observations & Results: <br> </li>
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<h2><b>V06_06 </b></h2><br>
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<b>Chemical transformartion of the <i>flhDC</i>-promoter constructs into <i>E. coli</i> (DH10B)</i></b><br>
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<ul>
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<li>Experiment: <br>
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For the chemical transformation the standard protocol was followed.
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<li>Observations & Results: <br>
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The transformation was successful since all plates showed numeours colonoes except the negative control.However, on the plastes with 18K and 18O few red colonies could be observed. These were again rejected.
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<h2><b>V06_07 </b></h2><br>
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<b>Preparation of over night cultures</i></b><br>
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<ul>
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<li>Experiment: <br>
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Over night cultures were preaped of three colonies of each constructs in order to isolate the plasmids.
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<h2><b><a name="week7#3">#3 Chemoreceptor Library</a></b></h2>
 
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<a href="https://2012.igem.org/Team:Goettingen/Notebook">Back to overview</a><br>
 
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<p align="justify" style="line-height:1.6em">
 
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Under process...
 
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<h2><b></b></h2><br>
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<h2><b>V06_08 </b></h2><br>
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<b></b><br>
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<b>V06_08_1 Miniprep of the new <i>flhDC</i>-promoter constructs</b><br>
<ul>
<ul>
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<li>Experiment:  <br> </li>
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<li>Experiment:  <br>
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<li>Description: <br> </li>
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Minipreps were conducted with PeqGOLD MiniPrep Kit (Peqlab) according to the user manual.
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<li>Observations & Results: <br> </li>
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<li>
</ul>
</ul>
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<b><i/></b><br>
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<b>V06_08_2  Test digestion of the new <i>flhDC</i>-promoter constructs</b><br>
<ul>
<ul>
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<li>Experiment: </li>
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<li>Experiment: <br>
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<li>Description: <br> </li>
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In order to prove correct insertion of <i>flhDC</i> a test digestion was performed using SpeI and XbaI according to the protocol.<br>  
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<li>Observations & Results: <br> </li>
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<li>Observations & Results: <br>
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</ul>
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For each construct two to three clones hosting the correctly inserted gene in the plasmid could be obtained. Those colonies still containing the rfp gene and were thus rejected.</li>
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Revision as of 09:49, 14 September 2012

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