Team:Goettingen/week9-2

From 2012.igem.org

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<h2><b>V06_25 </b></h2><br>
 
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<b>Chemical transformartion of the BioBrick plasmid pSB1C3 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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In order to gain further plasmid material of the new vector pSB1C3 was transformed into E. coli(DH10B) as described in the protocol. <br>
 
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<li>Observations & Results: <br>
 
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The transformation failed. Only the positive control featured colony formation. We found out that accidentally ampicillin containing plates were used whereas the plasmid obtains a chloramphenicol resistance. Thus, the out come is not surprising.
 
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<h2><b>V06_26 </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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Since we received a new <i>E. coli</i> (DH10B) strain, over night cultures were prepared in order to produce new competent cells.The following protocol was considered.<br>
 
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<li>Observations & Results: <br>
 
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The performance of a test transformation proved the functionality of the new competent cells.
 
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<br>
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<h2><b>V06_25 </b></h2><br>
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<b>V06_25_1 Chemical transformartion of the BioBrick plasmid pSB1C3 into <i>E. coli</i> (DH10B)</b><br>
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<h2><b>V06_19 </b></h2><br>
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<b>V06_19_1 Repetition of the chemical transformartion of the BioBrick plasmid pSB1C3 into <i>E. coli</i> (DH10B) </b><br>
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<ul>
<ul>
<li>Experiment: <br>
<li>Experiment: <br>
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Minipreps were conducted with PeqGOLD MiniPrep Kit (Peqlab) according to the user manual.</li>
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In order to gain further plasmid material of the new vector pSB1C3 was transformed into E. coli(DH10B) as described in the protocol. </li>
<li>Observations & Results: <br>
<li>Observations & Results: <br>
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The transformation was successful.Due to the possesion of a gene encoding for RFP the developed colonied featured a red color. On the negative control no colonied were observed.
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The transformation failed. Only the positive control featured colony formation. We found out that accidentally ampicillin containing plates were used whereas the plasmid obtains a chloramphenicol resistance. Thus, the out come is not surprising.
</li></ul>
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<b>V06_19_2 Repetition of the chemical transformartion of the <i>araC</i>-promoter into <i>E. coli</i> (DH10B) </b><br>
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<b>V06_25_2 Preparation of over night cultures </b><br>
<ul>
<ul>
<li>Experiment:  <br>
<li>Experiment:  <br>
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Since the last time the fact that the <i>araC</i>-promoter I0500 (14N) is inducible by ITPG was not considered, this time the substance was added to the LB-Medium (end concentration 100 µM). This is the only deogation from the standard transformation protocol. <br>
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Since we received a new <i>E. coli</i> (DH10B) strain, over night cultures were prepared in order to produce new competent cells.The following protocol was considered.</li>
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<li>Observations & Results: <br>
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The transformation was again not successful. Only the positive control showed any growth. </li>
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</ul><br>
</ul><br>

Revision as of 10:56, 15 September 2012

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