Team:Goettingen/week7-2

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                              <!--</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>
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                              </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> -->
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                              <li><a href="https://2012.igem.org/Team:Goettingen/Human_Practice/Panel_Discussion"><span><span>Panel Discussion</span></span></a></li>-->
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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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<h2><b><a name="week6#2">#2 Speed Improvement - 6th week</a></b></h2>
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<h2><b><a name="week7#2">#2 Speed Improvement - 7th week</a></b></h2>
<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>
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<h2><b>V06_04 </b></h2><br>
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<h2><b>V06_11 </b></h2><br>
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<b>Chemical retransformartion of the <i>flhDC</i>-promoter constructs into <i>E. coli</i> (DH10B)</i></b><br>
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<b>V06_11_1 Miniprep of the new <i>flhDC</i>-promoter constructs</b><br>
<ul>
<ul>
-
<li>Experiment: <br>  
+
<li>Experiment: <br>
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For the chemical retransformation the standard protocol for transformation was followed. The following constructs were transformed into
+
Minipreps were conducted with PeqGOLD MiniPrep Kit (Peqlab) according to the user manual.
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E. coli: <br>
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</ul>
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20E - #2 <br>
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<br>
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20E - #3 <br>
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<b>V06_11_2 Chemical retransformation of the new <i>flhDC</i>-promoter constructs into <i>E. coli</i> (DH10B)</b><br>
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18M - #2 <br>
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<ul>
-
18M - #3 <br>
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<li>Experiment:  <br>
-
18C - #1 <br>
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For the chemical retransformation the standard <a href="https://2012.igem.org/Team:Goettingen/Project/Methods">protocol</a> was followed. The following constructs were transformed into <i>E. coli</i>:<br>
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18C - #2 <br></li>  
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<br>
 +
20G - #1 <br>
 +
20G - #2 <br>
 +
2G - #1 <br>
 +
2G - #2 <br>
 +
2G - #3 <br>
 +
20I - #2 <br>
 +
20I - #3 <br>
 +
18O - #2 <br>
 +
18O - #3 <br>
 +
18K - #1 <br>
 +
18K - #2 <br>
 +
18K - #3 <br></li>
 +
<br>
<li>Observations & Results: <br>
<li>Observations & Results: <br>
-
The retransformation was successful since all plates showed numeours colonoes except the negative control.
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The retransformation was successful since all plates showed numeours colonies except the negative control.
-
<br> </li>
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<h2><b>V06_05 </b></h2><br>
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<h2><b>V06_12 </b></h2><br>
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<b>V06_05_1 Preparative double digestion of <i>flhDC</i> and plasmids containing further promoter</b><br>
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<b>Preparation of over night cultures</i></b><br>
<ul>
<ul>
-
<li>Experiment: <br>
+
<li>Experiment: <br>
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In order to clone <i>flhDC</i> behind further promoters, the purified PCR product was digested with XbaI and PstI, whereas the BioBrick plasmids were cut with SpeI and PstI. The digestion was performed as described in the protocol. This time the follwing promoters were used: <br>
+
In order to gain further plasmid material over night cultures of one colonie of all eight <i>flhDC</i>-promoter constructs were prepared for subsequent plasmid isolation. <br>
-
J23113 - 20G - x21 <br>
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<br>
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J23109  - 2G - x106 <br>
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- 20E - #2 <br>
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J23114 - 20I - x256 <br>
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- 20G - #1 <br>
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J23106 - 18O - x1185 <br>
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- 2G - #1 <br>
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J23104 - 18K - x 1831 <br>
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- 20I - #2 <br>
-
 
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- 18M - #2 <br>
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- 18O - #2 <br>
 +
- 18K - #1 <br>
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- 18C - #2 <br>
<br>
<br>
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<b>V06_05_2  Insertion of <i>flhDC</i> into the BioBrick plamids</b><br>
 
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<li>Experiment:  <br>
 
-
The ligation of the digested flhDC with the BioBrick plasmids was conducted as described in the protocol. <br> </li>
 
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<b>Chemical transformartion of the <i>flhDC</i>-promoter constructs into <i>E. coli</i> (DH10B)</i></b><br>
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<h2><b>V06_13 </b></h2><br>
 +
<b>V06_13_1 Miniprep of the new <i>flhDC</i>-promoter constructs</b><br>
<ul>
<ul>
-
<li>Experiment: <br>  
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<li>Experiment: <br>
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For the chemical transformation the standard protocol was followed.
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Minipreps were conducted with PeqGOLD MiniPrep Kit (Peqlab) according to the user manual.</li></ul>
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<br>
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<b>V06_13_2 Chemical transformation of pBAD-<i>sfGFP</i> into <i>E. coli</i> (DH10B)</b><br>
 +
<ul>
 +
<li>Experiment:  <br>
 +
For the chemical transformation the standard <a href="https://2012.igem.org/Team:Goettingen/Project/Methods">protocol</a> was followed. <br>
<li>Observations & Results: <br>
<li>Observations & Results: <br>
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The transformation was successful since all plates showed numeours colonoes except the negative control.However, on the plastes with 18K and 18O few red colonies could be observed. These were again rejected.
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The transformation was successful since all plates showed numeours colonies except the negative control.</li>
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</ul>
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</ul><br>
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<h2><b>V06_07 </b></h2><br>
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<h2><b>V06_14 </b></h2><br>
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<b>Preparation of over night cultures</i></b><br>
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<b> Preparation of over night cultures</i></i></b><br>
<ul>
<ul>
-
<li>Experiment: <br>  
+
<li>Experiment: <br>
-
Over night cultures were preaped of three colonies of each constructs in order to isolate the plasmids.
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Over night cultures of the pBAD-<i>sfGFP</i> clones were prepared in order to isolate the plasmids. <br>
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<br> </li>
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</ul>
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<h2><b>V06_08 </b></h2><br>
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<h2><b>V06_15 </b></h2><br>
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<b>V06_08_1 Miniprep of the new <i>flhDC</i>-promoter constructs</b><br>
+
<b> Miniprep of pBAD-<i>sfGFP</i></i></i></b><br>
<ul>
<ul>
-
<li>Experiment: <br>
+
<li>Experiment: <br>
-
Minipreps were conducted with PeqGOLD MiniPrep Kit (Peqlab) according to the user manual.  
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Minipreps were conducted with PeqGOLD MiniPrep Kit (Peqlab) according to the user manual. <br>
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<li>
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-
</ul>
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<br>
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<b>V06_08_2  Test digestion of the new <i>flhDC</i>-promoter constructs</b><br>
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<ul>
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<li>Experiment:  <br>
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In order to prove correct insertion of <i>flhDC</i> a test digestion was performed using SpeI and XbaI according to the protocol.<br>
+
-
<li>Observations & Results: <br>
+
-
For each construct two to three clones hosting the correctly inserted gene in the plasmid could be obtained. Those colonies still containing the rfp gene and were thus rejected.</li>
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<br></td></tr>
<br></td></tr>
</table>
</table>
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<br>
<br>
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<body id="top">
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<a href="https://2012.igem.org/Team:Goettingen/Notebook">Back to overview</a><br>
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<a href="#top">&uarr; Return to top</a>
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<b>Important pages</b>:<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>;
 
-
<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/Notebook">Notebook</a>;
 
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<a href="https://2012.igem.org/Team:Goettingen/Saftey">Saftey</a>;
 
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<a href="https://2012.igem.org/Team:Goettingen/Attributions">Attributions</a>
 
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Latest revision as of 17:17, 22 September 2012

Deutsch  / English 

#2 Speed Improvement - 7th week

Back to overview

V06_11


V06_11_1 Miniprep of the new flhDC-promoter constructs
  • Experiment:
    Minipreps were conducted with PeqGOLD MiniPrep Kit (Peqlab) according to the user manual.

V06_11_2 Chemical retransformation of the new flhDC-promoter constructs into E. coli (DH10B)
  • Experiment:
    For the chemical retransformation the standard protocol was followed. The following constructs were transformed into E. coli:

    20G - #1
    20G - #2
    2G - #1
    2G - #2
    2G - #3
    20I - #2
    20I - #3
    18O - #2
    18O - #3
    18K - #1
    18K - #2
    18K - #3

  • Observations & Results:
    The retransformation was successful since all plates showed numeours colonies except the negative control.


V06_12


Preparation of over night cultures
  • Experiment:
    In order to gain further plasmid material over night cultures of one colonie of all eight flhDC-promoter constructs were prepared for subsequent plasmid isolation.

    - 20E - #2
    - 20G - #1
    - 2G - #1
    - 20I - #2
    - 18M - #2
    - 18O - #2
    - 18K - #1
    - 18C - #2



V06_13


V06_13_1 Miniprep of the new flhDC-promoter constructs
  • Experiment:
    Minipreps were conducted with PeqGOLD MiniPrep Kit (Peqlab) according to the user manual.

V06_13_2 Chemical transformation of pBAD-sfGFP into E. coli (DH10B)
  • Experiment:
    For the chemical transformation the standard protocol was followed.
  • Observations & Results:
    The transformation was successful since all plates showed numeours colonies except the negative control.


V06_14


Preparation of over night cultures
  • Experiment:
    Over night cultures of the pBAD-sfGFP clones were prepared in order to isolate the plasmids.


V06_15


Miniprep of pBAD-sfGFP
  • Experiment:
    Minipreps were conducted with PeqGOLD MiniPrep Kit (Peqlab) according to the user manual.


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Retrieved from "http://2012.igem.org/Team:Goettingen/week7-2"