Team:Goettingen/week18-2

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<a name="week3#2">#2 Speed Improvement - 18th week</a>

<a href="https://2012.igem.org/Team:Goettingen/Notebook">Back to overview</a>

</table>

V08_27

V08_27_1 Miniprep of the tar-promoter constructs

Experiment:
The Minipreps were conducted using the PeqGOLD MiniPrep Kit (Peqlab) according to the user manual.

V08_27_2 Chemical retransformation of the tar-promoter constructs into E. coli BL21

Experiment:
The retransformation was performed as described in the standard protocol.
Observations & Results:
The retransformation was successful since numerous colonies could be obtained except for the empty negative control.

V08_27_3 Chemical transformation of different motility increasing constructs into E. coli MG1655

Experiment:
For we aimed to investigate the effect of our constructs on cells that are used to swim competent MG1655 cells were transformed with the following constructs:
fliC (DH10B)
fliC (Salmonella)
motA
motB
yhjH

18M-flhDC
18O-flhDC
18C-flhDC
Observations & Results:
The transformation was not successful. The next morning only one plate (the positive control) featured any visible colonies. However, after several additional hours of incubation also some very small colonies appeared at some other plates. Nevertheless, the efficiency was very low. We suggest that this is normal since MG1655 is no cloning strain and might have thus a much lower transformation efficiency. This idea was supported by our inquiries.

V08_27_4 Preparation of over night cultures

Experiment:
In oder to perform a Real-Time Quantitative Reverse Transcription PCR over night cultures of the tar-promoter constructs in BL21 were prepared.

</ul>

V08_28

Preparative double digestion of the flhDC-promoter constructs and pSB1C3

Experiment:
The flhDC-promoter constructs as well as the vector pSB1C3 still including tar were digested with EcoRI and PstI as described in the protocol. Afterwards, the fragments of the desired size were cut out and frozen in order to extract the DNA from the gel slices the following day.
Observations & Results:
The gel featured clearly visible bands of the expected length and thus the digestion was successful.

V08_29

V08_19_1 Purification of the the flhDC-promoter constructs and pSB1C3

Experiment:
The restriction product was purified using peqGOLD Gel Extraction Kit (Peqlab) according to the manual.

V08_29_2 Insertion of the flhDC-promoter constructs into pSB1C3

Experiment:
The ligation of the digested flhDC-promoter constructs with pSB1C3 was conducted as described in the protocol.

V08_30

Miniprep of the flhDC-promoter constructs in pUC18

Experiment:
In order to obtain further plasmid material Minipreps were conducted using the PeqGOLD MiniPrep Kit (Peqlab) according to the user manual.

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-                        <!--<li><li><a href="https://2012.igem.org/Team:Goettingen/groups#library"><span><span><div style="text-indent:20px;">&#8901; Library</div></span></span></a></li>             -->
-                        <li><a href="https://2012.igem.org/Team:Goettingen/Seminars"><span><span>Seminars</span></span></a></li>
-                        <!--<li><li><a href="https://2012.igem.org/Team:Goettingen/seminars#announce"><span><span><div style="text-indent:20px;">&#8901; Announcements</div></span></span></a></li>    -->
-                        <!--<li><li><a href="https://2012.igem.org/Team:Goettingen/seminars#presentations"><span><span><div style="text-indent:20px;">&#8901; Past Presentations</div></span></span></a></li> -->
-                        <li><a href="https://2012.igem.org/Team:Goettingen/Work_Impressions"><span><span>Work Impressions</span></span></a></li>
-                        </ul>
-                        <!--[if lte IE 6]></td></tr></table></a><![endif]-->
-                </li>
-                <li>
-                        <a href="https://2012.igem.org/Team:Goettingen/Project" style="color: white;">Project<!--[if gt IE 6]><!--></a><!--<![endif]-->
-                        <!--[if lt IE 7]><table border="0" cellpadding="0" cellspacing="0"><tr><td><![endif]-->
-                        <ul class="DropDownMenu" id="MB1-DDM2-SM3">
-                        <li><a href="https://2012.igem.org/Team:Goettingen/Project"><span><span>Our Project</span></span></a></li>
-                        <!--<li></li><li><a href="https://2012.igem.org/Team:Goettingen/Chemotaxis"><span><span><div style="text-indent:20px;">&#8901; Chemotaxis</div></span></span></a></li>  -->
-                        <!--<li></li><li><a href="https://2012.igem.org/Team:Goettingen/Poster"><span><span><div style="text-indent:20px;">&#8901; Poster</div></span></span></a></li>    -->
-                        <li><a href="https://2012.igem.org/Team:Goettingen/Project/Materials"><span><span>Materials</span></span></a>
-                        <li><a href="https://2012.igem.org/Team:Goettingen/Project/Methods"><span><span>Methods</span></span></a>
-                        <li><a href="https://2012.igem.org/Team:Goettingen/Project/Computational_Data"><span><span>Computational Data</span></span></a>
-                        <!--</li><li><a href="https://2012.igem.org/Team:Goettingen/Maps"><span><span><div style="text-indent:20px;">&#8901; Maps</div></span></span></a></li>
-                        </li><li><a href="https://2012.igem.org/Team:Goettingen/Sequences"><span><span><div style="text-indent:20px;">&#8901; Sequences</div></span></span></a></li> -->
-                        <li><a href="https://2012.igem.org/Team:Goettingen/Project/Bioinformatical_Tool"><span><span>Bioinformatical Tools</span></span></a>
-                        <li><a href="https://2012.igem.org/Team:Goettingen/Project/Lab_Work_Flow"><span><span>Lab Work Flow</span></span></a>
-                        <!--</li><li><a href="https://2012.igem.org/Team:Goettingen/Maps"><span><span><div style="text-indent:20px;">&#8901; Golden Lab Rules</div></span></span></a></li>
-                        </li><li><a href="https://2012.igem.org/Team:Goettingen/Sequences"><span><span><div style="text-indent:20px;">&#8901; Daily Check List</div></span></span></a></li> -->
-                        </ul>
-                        <!--[if lte IE 6]></td></tr></table></a><![endif]-->
-                </li>
-                <li>
-                        <a href="https://2012.igem.org/Team:Goettingen/Notebook" style="color: white;">Notebook<!--[if gt IE 6]><!--></a><!--<![endif]-->
-                        <!--[if lt IE 7]><table border="0" cellpadding="0" cellspacing="0"><tr><td><![endif]-->
-                                <ul class="DropDownMenu" id="MB1-DDM2">
-                                <li><a href="https://2012.igem.org/Team:Goettingen/Notebook"><span><span>Overview</span></span></a></li>
-                                <li><a href="https://2012.igem.org/Team:Goettingen/Notebook/Summary"><span><span>Summary</span></span></a></li>
-                                <li><a href="https://2012.igem.org/Team:Goettingen/Notebook/Discussion"><span><span>Discussion</span></span></a></li>
-                                </ul>
-                                <!--[if lte IE 6]></td></tr></table></a><![endif]-->
-                </li>
-                <li>
-                        <a href="https://2012.igem.org/Team:Goettingen/iGEM" style="color: white;">iGEM<!--[if gt IE 6]><!--></a><!--<![endif]-->
-                        <!--[if lt IE 7]><table border="0" cellpadding="0" cellspacing="0"><tr><td><![endif]-->
-                                <ul class="DropDownMenu" id="MB1-DDM6">
-                                <li><a href="https://2012.igem.org/Team:Goettingen/iGEM"><span><span>What is iGEM?</span></span></a></li>
-                                <!--<li><a href="https://2012.igem.org/Team:Goettingen/BioBrick_System"><span><span>iGEM BioBrick System</span></span></a></li>
-                                <li><a href="https://2012.igem.org/Team:Goettingen/Part_Registry"><span><span>iGEM Part Registry</span></span></a></li>  -->
-                                <li><a href="https://2012.igem.org/Team:Goettingen/iGEM/Parts_Submitted"><span><span>Parts Submitted</span></span></a></li>
-                                <li><a href="https://2012.igem.org/Team:Goettingen/iGEM/Attributions"><span><span>Attributions</span></span></a></li>
-                                <li><a href="https://2012.igem.org/Team:Goettingen/iGEM/Saftey"><span><span>Saftey</span></span></a></li>>
-                                </ul>
-                                <!--[if lte IE 6]></td></tr></table></a><![endif]-->
-                </li>
-                <li>
-                        <a href="https://2012.igem.org/Team:Goettingen/Goettingen" style="color: white;">G&ouml;ttingen<!--[if gt IE 6]><!--></a><!--<![endif]-->
-                        <!--[if lt IE 7]><table border="0" cellpadding="0" cellspacing="0"><tr><td><![endif]-->
-                                <ul class="DropDownMenu" id="MB1-DDM5">
-                                <li><a href="https://2012.igem.org/Team:Goettingen/Goettingen"><span><span>City</span></span></a></li>
-                                <li><a href="https://2012.igem.org/Team:Goettingen/Goettingen/University"><span><span>Georg-August-University</span></span></a></li>
-                                <li><a href="https://2012.igem.org/Team:Goettingen/Goettingen/MPI"><span><span>Max-Planck-Institute</span></span></a></li>
-                                <li><a href="https://2012.igem.org/Team:Goettingen/Goettingen/S1-Demo_Lab"><span><span>S1-Demo Lab</span></span></a></li>
-                        <!--[if lt IE 7]><table border="0" cellpadding="0" cellspacing="0"><tr><td><![endif]-->
-                                        <!--[if lte IE 6]></td></tr></table></a><![endif]-->
-                                </li>
-                                </ul>
-                                <!--[if lte IE 6]></td></tr></table></a><![endif]-->
-                </li>
-                <li style="width: 160px">
-                        <a href="https://2012.igem.org/Team:Goettingen/Human_Practice" style="color: white;">Human Practice<!--[if gt IE 6]><!--></a><!--<![endif]-->
-                        <!--[if lt IE 7]><table border="0" cellpadding="0" cellspacing="0"><tr><td><![endif]-->
-                                <ul class="DropDownMenu" id="MB1-DDM5">
-                               <li><a href="https://2012.igem.org/Team:Goettingen/Human_Practice"><span><span>Why Human Practice?</span></span></a></li>
-                               <li><a href="https://2012.igem.org/Team:Goettingen/Human_Practice/Public_and_Media"><span><span>Public and Media</span></span></a></li>
-                               <!--</li><li><a href="https://2012.igem.org/Team:Goettingen/Newspaper"><span><span><div style="text-indent:20px;">&#8901; Newspaper</div></span></span></a></li>
-                               </li><li><a href="https://2012.igem.org/Team:Goettingen/Conference"><span><span><div style="text-indent:20px;">&#8901; Conference</div></span></span></a></li>
-                               </li><li><a href="https://2012.igem.org/Team:Goettingen/Synthetic_Biology_Day"><span><span><div style="text-indent:20px;">&#8901; Synthetic Biology Day</div></span></span></a></li> -->
-                               <li><a href="https://2012.igem.org/Team:Goettingen/Human_Practice/Survey"><span><span>Survey</span></span></a></li>
-                               <li><a href="https://2012.igem.org/Team:Goettingen/Human_Practice/Panel_Discussion"><span><span>Panel Discussion</span></span></a></li>-->
-                               <li><a href="https://2012.igem.org/Team:Goettingen/Human_Practice/Flash_coli"><span><span>Flash Coli</span></span></a></li>
-                               </ul>
-                                <!--[if lte IE 6]></td></tr></table></a><![endif]-->
-                </li>
-                <li>
-                        <a href="https://2012.igem.org/Team:Goettingen/Sponsoring" style="color: white;">Sponsoring</a>
-                        <!--[if lt IE 7]><table border="0" cellpadding="0" cellspacing="0"><tr><td><![endif]-->
-                                <ul class="DropDownMenu" id="MB1-DDM2-SM3">
-                               <li><a href="https://2012.igem.org/Team:Goettingen/Sponsoring"><span><span>Interested in...?</span></span></a></li>
-                                <!-- <li><a href="https://2012.igem.org/Team:Goettingen/Human_Practice/Sponsors"><span><span>Sponsors</span></span></a></li>
-                                <li><a href="https://2012.igem.org/Team:Goettingen/Human_Practice/Supporters"><span><span>Supporters</span></span></a></li>     -->
-                </li>
-        </ul>
-</div>
-<table>
-                                 <!-- Text body -->
-<tbody><tr valign="top" align="left">
-<td style="padding: 0pt 20px 0pt 0pt;" width="650px">
-<font face="Verdana" size="-1">
-<br>
-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>
@@ Line 624: / Line 111: @@
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-<br>
-<body id="top">
-<a href="#top">&uarr; Return to top</a>
-<br>
 <br>
 <a href="https://2012.igem.org/Team:Goettingen/Notebook">Back to overview</a><br>
 <br>
 </td>
-<table bordercolor="black" border="1 px" width="600 px"><tr><td>
-<b>Important pages</b>:<br>
-<a href="https://2012.igem.org/Team:Goettingen">Home</a>;
-<a href="https://2012.igem.org/Team:Goettingen/Team">Team</a>;
-<a href="https://igem.org/Team.cgi?year=2012">Official Team Profile</a>;
-<a href="https://2012.igem.org/Team:Goettingen/Project">Project</a>;
-<a href="https://2012.igem.org/Team:Goettingen/Parts">Parts submitted to the Registry</a>;
-<a href="https://2012.igem.org/Team:Goettingen/Modeling">Modeling</a>;
-<a href="https://2012.igem.org/Team:Goettingen/Notebook">Notebook</a>;
-<a href="https://2012.igem.org/Team:Goettingen/Saftey">Saftey</a>;
-<a href="https://2012.igem.org/Team:Goettingen/Attributions">Attributions</a>
-</td></tr>
-</table>
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