Team:Goettingen/week6-2

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#2 Speed Improvement - 6th week

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V06_04

Chemical transformartion of the flhDC-promoter constructs into E. coli (DH10B)

Experiment:
Over night cultures for the preparation of new competent cells were prepared using E. coli (DH10B) cells without any resistance.

V05_25

V05_25_1 Preparing chemically competent cells of E. coli (DH10B)

Experiment:
The preparation of the competent cells was performed according to the following protocol. This time cells without any resistance were utilized for inoculation

V05_25_2 Chemical transformation of competent E. coli (DH10B)

Experiment:
For the chemical transformation the standard protocol was followed. The following constructs were transformed into
E. coli
J23112 - 20E - x1
J23113 - 20G - x21
J23109 - 2G - x106
J23114 - 20I - x256
J23105 - 18M - x623
J23106 - 18O - x1185
J23104 - 18K - x 1831
J23100 - 18 C - x 2547
I0500 - 14N - pBAD/araC promoter
Observations & Results:
The transformation was successful. Numerous colonies on all plates as well as on the positive control. Now growth could be observed on the negative control plate, so the competent cells are preparation of the competent cells was correct this time. Additionally, no growth could be observed at the 14N plate. This is the only vector containing a kanamycin resistance instead of ampicillin. Eventually, the kanamycin concentration was too strong and inhibited growth.
The strain (DH10B) grows very fast, thus try not to incubate too long to prevent formation of satellite colonies.
The cells hosting plasmids with strong promoters are colored red due to the rfp gene fused to the promoter. This phenomenon can be used to verify correct transformation.

V05_27

Preparation of over night cultures

Experiment:
In order to isolate the plasmids, over night cultures of the E. coli (DH10B) cells transformed with the different constitutive promoters were prepared.

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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>
 <br>
-<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>
-<ul>
-<li class="toclevel-1"><a href="#week6"><span class="tocnumber">1</span> <span class="toctext">6th week (04th to 10th June)</span></a></li>
-<li class="toclevel-1"> <div style="text-indent:10px;"><a href="#week6#1"><span class="tocnumber"></span> <span class="toctext">#1 Selection / Swimming</span></a></li>
-<li class="toclevel-1"> <div style="text-indent:10px;"><a href="#week6#2"><span class="tocnumber"></span> <span class="toctext">#2 Speed Improvement</span></a></li>
-<li class="toclevel-1"> <div style="text-indent:10px;"><a href="#week6#3"><span class="tocnumber"></span> <span class="toctext">#3 Chemoreceptor Library</span></a></li>
-</ul>
-</td></tr></tbody></table>
-<br>
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 <font face="Verdana" size="-1">
-<h2><b><a name="week6#1">#1 Selection / Swimming</a></b></h2>
+<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>
 <br>
 <p align="justify" style="line-height:1.6em">
-Under process..
 </p>
 <table cellpadding="20 px" border="1" bordercolor="black" valign="top">
 <tr bordercolor="black" valign="top">
 <td width="900" bordercolor="black" valign="top">
-<h2><b></b></h2><br>
+<h2><b>V06_04 </b></h2><br>
-<b></b><br>
+<b>Chemical transformartion of the <i>flhDC</i>-promoter constructs into <i>E. coli</i> (DH10B)</i></b><br>
 <ul>
-<li>Experiment:  <br> </li>
+<li>Experiment: <br>
-<li>Description: <br> </li>
+Over night cultures for the preparation of new competent cells were prepared using <i>E. coli</i> (DH10B) cells without any resistance. </li>
-<li>Observations & Results: <br> </li>
+<br> </li>
-</ul>
-<br>
-<b><i/></b><br>
-<ul>
-<li>Experiment: </li>
-<li>Description: <br> </li>
-<li>Observations & Results: <br> </li>
 </ul>
 <br></td></tr>
 </table>
-<br>
-<br>
-<br>
-<br>
-<font face="Verdana" size="-1">
-<h2><b><a name="week6#2">#2 Speed Improvement</a></b></h2>
-<a href="https://2012.igem.org/Team:Goettingen/Notebook">Back to overview</a><br>
 <br>
 <p align="justify" style="line-height:1.6em">
-Under process...
 </p>
 <table cellpadding="20 px" border="1" bordercolor="black" valign="top">
 <tr bordercolor="black" valign="top">
 <td width="900" bordercolor="black" valign="top">
-<h2><b></b></h2><br>
+<h2><b>V05_25 </b></h2><br>
-<b></b><br>
+<b>V05_25_1 Preparing chemically competent cells of <i>E. coli</i> (DH10B)</b><br>
 <ul>
-<li>Experiment:  <br> </li>
+<li>Experiment:  <br>
-<li>Description: <br> </li>
+The preparation of the competent cells was performed according to the following protocol. This time cells without any resistance were utilized for inoculation</li>
-<li>Observations & Results: <br> </li>
 </ul>
 <br>
-<b><i/></b><br>
+<b>V05_25_2 Chemical transformation of competent <i>E. coli</i> (DH10B) </b><br>
 <ul>
-<li>Experiment: </li>
+<li>Experiment:  <br>
-<li>Description: <br> </li>
+For the chemical transformation the standard protocol was followed. The following constructs were transformed into <br>
-<li>Observations & Results: <br> </li>
+ <i>E. coli</i> <br>
+J23112 - 20E - x1 <br>
+J23113 - 20G - x21 <br>
+J23109  - 2G - x106 <br>
+J23114 - 20I - x256 <br>
+J23105 - 18M - x623 <br>
+J23106 - 18O - x1185 <br>
+J23104 - 18K - x 1831 <br>
+J23100 - 18 C - x 2547 <br>
+I0500 - 14N - pBAD/araC promoter </li>
+<li>Observations & Results: <br>
+The transformation was successful. Numerous colonies on all plates as well as on the positive control. Now growth could be observed on the negative control plate, so the competent cells are preparation of the competent cells was correct this time. Additionally, no growth could be observed at the 14N plate. This is the only vector containing a kanamycin resistance instead of ampicillin. Eventually, the kanamycin concentration was too strong and inhibited growth.<br>
+The strain (DH10B) grows very fast, thus try not to incubate too long to prevent formation of satellite colonies. <br>
+The cells hosting plasmids with strong promoters are colored red due to the rfp gene fused to the promoter. This phenomenon can be used to verify correct transformation.<br> </li>
 </ul>
 <br></td></tr>
 </table>
-<br>
-<br>
 <br>
-<font face="Verdana" size="-1">
-<h2><b><a name="week6#3">#3 Chemoreceptor Library</a></b></h2>
-<a href="https://2012.igem.org/Team:Goettingen/Notebook">Back to overview</a><br>
-<br>
-<p align="justify" style="line-height:1.6em">
-Under process...
-</p>
 <table cellpadding="20 px" border="1" bordercolor="black" valign="top">
 <tr bordercolor="black" valign="top">
 <td width="900" bordercolor="black" valign="top">
-<h2><b></b></h2><br>
+<h2><b>V05_27 </b></h2><br>
-<b></b><br>
+<b>Preparation of over night cultures</i></b><br>
 <ul>
-<li>Experiment:  <br> </li>
+<li>Experiment: <br>
-<li>Description: <br> </li>
+In order to isolate the plasmids, over night cultures of the <i>E. coli</i> (DH10B) cells transformed with the different constitutive promoters were prepared.</li>
-<li>Observations & Results: <br> </li>
+<br> </li>
-</ul>
-<br>
-<b><i/></b><br>
-<ul>
-<li>Experiment: </li>
-<li>Description: <br> </li>
-<li>Observations & Results: <br> </li>
 </ul>
 <br></td></tr>
 </table>
-<br>
-<br>
-<br>
-<br>
-<br>
-<br>
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