Team:Goettingen/week20-2

From 2012.igem.org

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<h2><b>V09_10 </b></h2><br>
<h2><b>V09_10 </b></h2><br>
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<b>V09_10_1 Mini Prep of Psb1c3 (including motA, MotB or yhjH)</b><br>
+
<b>V09_10_1 Miniprep of pSB1C3 (including <i>motA</i>, <i>motB</i> or <i>yhjH</i>)</b><br>
<ul>
<ul>
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<li>Experiment: <br>Plasmids were prepped using PeqGOLD MiniPrep Kit (Peqlab)</li>
+
<li>Experiment: <br>Plasmids were prepped using PeqGOLD MiniPrep Kit (Peqlab) accirding to the manual.</li>
</ul>
</ul>
<br>
<br>
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<b>V09_10_2 Test digestion of Psb1c3 (including motA, motB or yhjH)</b><br>
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<b>V09_10_2 Test digestion of pSB1C3 (including <i>motA</i>, <i>motB</i> or <i>yhjH</i>)</b><br>
<ul>
<ul>
-
<li>Experiment: <br>Test digestion of Psb1c3 (including motA, yhjH or motB) with the restriction enzymes EcoRI and PstI. Test digest worked for all clones.</li>
+
<li>Experiment: <br>Test digestion of pSB1C3 (including <i>motA</i>, <i>motB</i> or <i>yhjH</i>) with the restriction enzymes EcoRI and PstI. Afterwards, the product was loaded in a 1% analytical gel in order to verify its success.<br> </li>
 +
<li>Observations & Results: <br>
 +
The digestion worked for all clones since fragments with the appropriate size were produced over all. </li>
</ul>
</ul>
<br>
<br>
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<b>V09_10_3 Digestion of motA, motB, yhjH and the promoters 20E, 20I and 18C</b><br>
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<b>V09_10_3 Digestion of <i>motA</i>, <i>motB</i>, <i>yhjH</i>and the promoters 20E, 20I and 18C</b><br>
<ul>
<ul>
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<li>Experiment: <br> MotA, MotB and yhjH were digested with XbaI and PstI. The plasmids with the different promoters were treated with SpeI and PstI. Digestion was controlled via gel-electrophoresis.The expected bands were isolated from the gel and purified via PeqGOLD Gelextraction Kit (Peqlab).</li>
+
<li>Experiment: <br> <i>motA</i>, <i>motB</i> or <i>yhjH</i> were digested with XbaI and PstI. The plasmids with the different promoters were treated with SpeI and PstI. The digestion was then controlled via gel-electrophoresis.<br> </li>
 +
<li>Observations & Results: <br>
 +
Since for all clones band of the expected size could be obtained these were isolated from the gel and purified using PeqGOLD Gelextraction Kit (Peqlab) according to the manual.</li>
</ul>
</ul>
<br>
<br>
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<b> V09_10_4 Quickchange of FliC</b><br>
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<b> V09_10_4 Quickchange of <i>fliC</i></b><br>
<ul>
<ul>
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<li>Experiment: <br> Quick Change overlapping PCR was performed using designed primers and Pfu Turbo polymerase (see Quick Change protocol in "protocols").No PCR product was observed</li>
+
<li>Experiment: <br> Quick Change overlapping PCR was performed using designed primers and Pfu Turbo polymerase (see Quick Change protocol in "protocols"). <br></li>
 +
<li>Observations & Results: <br>
 +
Once again the QC PCR failed for no PCR product was observed.</li>
</ul>
</ul>
<br></td></tr>
<br></td></tr>
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<h2><b>V09_11 </b></h2><br>
<h2><b>V09_11 </b></h2><br>
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<b>V09_11_1 Ligation and Transformation of motA, motB and yhjH</b><br>
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<b>V09_11_1 Ligation and Transformation of <i>motA</i>, <i>motB</i> and <i>yhjH</i></b><br>
<ul>
<ul>
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<li>Experiment: <br>  After the ligation of motA, motB and yhjH into the plasmid J61002 with different promoters (20E, 20I and 18C), the ligation products were transformed into the E. coli strain DH10B.</li>
+
<li>Experiment: <br>  After the ligation of <i>motA</i>, <i>motB</i> and <i>yhjH</i> into the plasmid J61002 with different promoters (20E, 20I and 18C), the ligation products were transformed into the <i>E. coli</i> strain DH10B.</li>
</ul>
</ul>
<br>
<br>
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<b>V09_11_2 Quickchange of FliC part 1</b><br>
+
<b>V09_11_2 Quickchange of <i>fliC</i> part 1</b><br>
<ul>
<ul>
<li>Experiment: <br> TEXT
<li>Experiment: <br> TEXT
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</ul>
</ul>
<br>
<br>
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<b>V09_12_2 Restriction and Ligation of promoter-RFP constructs into Psb1c3</b><br>
+
<b>V09_12_2 Restriction and Ligation of promoter-RFP constructs into pSB1C3</b><br>
<ul>
<ul>
<li>Experiment: <br>  Text</li>
<li>Experiment: <br>  Text</li>

Revision as of 13:37, 16 September 2012