Team:Goettingen/week6-3

From 2012.igem.org

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<b>Preparation of pUC18_TAR_QC</b><br>
<b>Preparation of pUC18_TAR_QC</b><br>
<ul>
<ul>
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<li>Experiment: <br>1% agarose gel was performed to analyze <i>Dpn</i>I digest. Gel extraction was performed as described before.
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<li>Experiment: <br>1% agarose gel was performed to analyze <i>Dpn</i>I digest. Gel extraction was performed as described before with PeqGOLD Gel extraction kit (Peqlab). Samples were stored at -20 °C.
</li>
</li>
</ul>
</ul>
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<h2><b>V06_07 </b></h2><br>
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<h2><b>V06_07-1 </b></h2><br>
<b>Preparation of chemocompetent cells of &Delta;<i>tar</i></b><br>
<b>Preparation of chemocompetent cells of &Delta;<i>tar</i></b><br>
<ul>
<ul>
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<li>Experiment: <br>Chemocompetent cells were prepared as described in <a href="https://2012.igem.org/Team:Goettingen/Project/Methods#Competent_Cells">protocol</a>.
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<li>Experiment: <br>Chemocompetent cells were prepared as described in this<a href="https://2012.igem.org/Team:Goettingen/Project/Methods#Competent_Cells">protocol</a>. ON cultures from 06.06. were used.
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</li>
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</ul>
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<h2><b>V06_07-2 </b></h2><br>
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<b>Miniprep of pUC18_TAR_QC</b><br>
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<ul>
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<li>Experiment: <br>Minipreps were done with PeqGOLD MiniPrep Kit (Peqlab) according to the user manual. A test digestion was performed using <i>Xba</i>I and the expected fragment of 4342 bp was observed on the corresponding gel. Minipreps were stored at -20 °C.
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</li>
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</ul>
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<br></td></tr>
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</table>
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<br>
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<h2><b>V06_08-1 </b></h2><br>
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<b>Preparative double digest of pUC18_TAR_QC</b><br>
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<ul>
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<li>Experiment: <br>Digest was performd with <i>Xba</i>I and <i>Pst</i>I (according to BioBrick standard) according to <a href="https://2012.igem.org/Team:Goettingen/Project/Methods">protocol</a>. A gelelectrophoresis of the digest was performed which showed the expected bands at 2662 bp and 1680 bp. The insert TAR_QC (1680 bp) was cut from the gel and a gel extraction was performed as described before. Stored at -20 °C.
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</li>
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</ul>
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<br></td></tr>
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</table>
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<br>
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<table cellpadding="20 px" border="1" bordercolor="black" valign="top">
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<tr bordercolor="black" valign="top">
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<td width="900" bordercolor="black" valign="top">
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<h2><b>V06_08-2 </b></h2><br>
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<b>Sequencing of pUC18_TAR_QC</b><br>
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<ul>
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<li>Experiment: <br>Sequencing reactions were done by Prof. Daniels Laboratory (University of Göttingen), Göttingen Genomics Lab and set up according to their <a href="https://2012.igem.org/Team:Goettingen/Project/Methods">protocol</a>. Primers used: pUC18_M13 for, pUC18_M13 rev, pUC18_TAR_Seqfor01, pUC18_TAR_Seqfor02. Dilutions 100 &mu;M, 10 &mu;M, 5 pmol.
</li>
</li>
</ul>
</ul>

Revision as of 15:21, 12 September 2012

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