Team:Goettingen/week6-3

From 2012.igem.org

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/*...............................................................................................*/
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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>
 
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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.
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<h2><b>V06_07 </b></h2><br>
<h2><b>V06_07 </b></h2><br>
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<b>Preparing chemocompetent cells of &Delta;''tar''</b><br>
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<b>V06_07_1 Preparation of chemocompetent cells of &Delta;<i>tar</i></b><br>
<ul>
<ul>
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<li>Experiment: <br>Preparation of overnight cultures for Miniprep of pUC18_TAR_QC.
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<li>Experiment: <br>
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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. </li>
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 +
<br>
 +
<b>V06_07_2 Miniprep of pUC18_TAR_QC </b><br>
 +
<ul>
 +
<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. </li>
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<h2><b>V06_08 </b></h2><br>
 +
<b>V06_08_1 Preparative double digest of pUC18_TAR_QC</b><br>
 +
<ul>
 +
<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.
 +
</li>
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<br>
<br>
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<b>V06_08_2 Sequencing of pUC18_TAR_QC</b><br>
 +
<ul>
 +
<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.
 +
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 +
<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>
<br>
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<a href="https://2012.igem.org/Team:Goettingen">Home</a>;
 
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<a href="https://2012.igem.org/Team:Goettingen/Team">Team</a>;
 
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<a href="https://igem.org/Team.cgi?year=2012">Official Team Profile</a>;
 
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<a href="https://2012.igem.org/Team:Goettingen/Project">Project</a>;
 
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<a href="https://2012.igem.org/Team:Goettingen/Parts">Parts submitted to the Registry</a>;
 
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<a href="https://2012.igem.org/Team:Goettingen/Modeling">Modeling</a>;
 
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<a href="https://2012.igem.org/Team:Goettingen/Saftey">Saftey</a>;
 
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{{GoettingenFooter}}

Latest revision as of 11:25, 22 September 2012

Deutsch  / English 

#3 Chemoreceptor Library - 6th Week

Back to overview

V06_04


Preparation of pUC18_TAR_QC
  • Experiment:
    1% agarose gel was performed to analyze DpnI digest. Gel extraction was performed as described before with peqGOLD Gel extraction kit (Peqlab). Samples were stored at -20 °C.


V06_05


Transforamtion of DH10B with pUC18_TAR_QC
  • Experiment:
    Transformation was performed according to protocol.


V06_06


Preparation of Miniprep cultures
  • Experiment:
    Preparation of overnight cultures for Miniprep of pUC18_TAR_QC.


V06_07


V06_07_1 Preparation of chemocompetent cells of Δtar
  • Experiment:
    Chemocompetent cells were prepared as described in this protocol. ON cultures from 06.06. were used.

V06_07_2 Miniprep of pUC18_TAR_QC
  • Experiment:
    Minipreps were done with peqGOLD Miniprep Kit (Peqlab) according to the user manual. A test digestion was performed using XbaI and the expected fragment of 4342 bp was observed on the corresponding gel. Minipreps were stored at -20 °C.


V06_08


V06_08_1 Preparative double digest of pUC18_TAR_QC
  • Experiment:
    Digest was performd with XbaI and PstI (according to BioBrick standard) according to protocol. 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.

V06_08_2 Sequencing of pUC18_TAR_QC
  • Experiment:
    Sequencing reactions were done by Prof. Daniels Laboratory (University of Göttingen), Göttingen Genomics Lab and set up according to their protocol. Primers used: pUC18_M13 for, pUC18_M13 rev, pUC18_TAR_Seqfor01, pUC18_TAR_Seqfor02. Dilutions 100 μM, 10 μM, 5 pmol.


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