Team:Braunschweig/Notebook

From 2013.igem.org

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     <h2><a href="#Week14">Week 14: August 18 - August 24, 2013</a></h2>
     <h2><a href="#Week14">Week 14: August 18 - August 24, 2013</a></h2>
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<img alt="Conti cultivation" src="https://2013.igem.org/File:Braunschweig_Lab_Week14_continuous_cultivation.jpg" height="200px" align="left" vspace="0" hspace="10" style="margin-right:5px"/>
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<img alt="Chromoprotein expression" src="https://static.igem.org/mediawiki/2013/f/fb/Braunschweig_Lab_expressed_chromoproteins.png" height="140px" align="left" vspace="0" hspace="10" style="margin-right:5px"/>
The focus of this week’s work was the preparation of the final constructs in order to take pictures under the fluorescence microscope. Furthermore we started the second continuous bioreactor culture.
The focus of this week’s work was the preparation of the final constructs in order to take pictures under the fluorescence microscope. Furthermore we started the second continuous bioreactor culture.
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Furthermore we miniprepped the final Plas contruct in <i>E. coli</i> JM109 and revised the sequencing results of the final eforRed-construct in <i>E. coli</i> Top10 F’. The final Prhl construct was sequence verified.<br>
Furthermore we miniprepped the final Plas contruct in <i>E. coli</i> JM109 and revised the sequencing results of the final eforRed-construct in <i>E. coli</i> Top10 F’. The final Prhl construct was sequence verified.<br>
We also conducted regulated and unregulated continuous bioreactor cultivations of mixed cultures of <i>E. coli</i> TOP10F'bearing final Prhl construct and <i>E. coli</i> JM109 containing the final Plas construct. The cultures were inoculated with a 70 % : 30 % ratio of <i>E. coli</i> Top10F' (Prhl construct)/<i>E. coli</i>(Plas construct) and samples were taken every hour. In order to determine the ratios of the strains during the cultivation, the samples were spread out on agar plates and incubated at 37°C. The cultivation was stopped after 25 h. The colonies on the plates were counted as shown below.<p>
We also conducted regulated and unregulated continuous bioreactor cultivations of mixed cultures of <i>E. coli</i> TOP10F'bearing final Prhl construct and <i>E. coli</i> JM109 containing the final Plas construct. The cultures were inoculated with a 70 % : 30 % ratio of <i>E. coli</i> Top10F' (Prhl construct)/<i>E. coli</i>(Plas construct) and samples were taken every hour. In order to determine the ratios of the strains during the cultivation, the samples were spread out on agar plates and incubated at 37°C. The cultivation was stopped after 25 h. The colonies on the plates were counted as shown below.<p>
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<p><img alt="August 19" src="https://static.igem.org/mediawiki/2013/a/ae/Braunschweig_Lab_Journal_August_19.png" width="340" vspace="20" align="left"/><img alt="August 19" src="https://static.igem.org/mediawiki/2013/e/e6/Braunschweig_Lab_Journal_August_19_2.png" width="400" vspace="20" align="left"/></p>
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<p><img alt="August 19" src="https://static.igem.org/mediawiki/2013/a/ae/Braunschweig_Lab_Journal_August_19.png" width="380" vspace="20" align="left"/><img alt="August 19" src="https://static.igem.org/mediawiki/2013/e/e6/Braunschweig_Lab_Journal_August_19_2.png" width="400" vspace="20" align="left"/></p>
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Furthermore 10 mM stock solutions of the homoserine lactones (autoinducers) of the rhl and las quorum sensing system (1.7 mg N-butyryl  homoserine lactone + 1 ml DMSO and 2.8 mg N-3-oxododecanoyl  homoserine lactone +  933 µl DMSO, respectively) were prepared.<br>
Furthermore 10 mM stock solutions of the homoserine lactones (autoinducers) of the rhl and las quorum sensing system (1.7 mg N-butyryl  homoserine lactone + 1 ml DMSO and 2.8 mg N-3-oxododecanoyl  homoserine lactone +  933 µl DMSO, respectively) were prepared.<br>
Additionally we repeated the continuous bioreactor cultivation we conducted Monday. Counting the colonies resulted in the diagrams shown below.<br>
Additionally we repeated the continuous bioreactor cultivation we conducted Monday. Counting the colonies resulted in the diagrams shown below.<br>
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<img alt="August 22" src="https://static.igem.org/mediawiki/2013/8/86/Braunschweig_Lab_Journal_August_22_2.png" width="400" vspace="20" align="left"/><img alt="August 22" src="https://static.igem.org/mediawiki/2013/1/17/Braunschweig_Lab_Journal_August_22_3.png" width="400" vspace="20" align="left"/>
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<img alt="August 22" src="https://static.igem.org/mediawiki/2013/8/86/Braunschweig_Lab_Journal_August_22_2.png" width="420" vspace="20" align="left"/><img alt="August 22" src="https://static.igem.org/mediawiki/2013/1/17/Braunschweig_Lab_Journal_August_22_3.png" width="400" vspace="20" align="left"/>
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Revision as of 00:03, 5 October 2013

Labjournal

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Braunschweig Labbook This is the documentation of our lab work. Achievements of each week are summerized followed by a daily discription of our experiments.

An overview on how we approached this project can be found under Approach. For detailed protocols of certain procedures please refer to Protocols. Attributions are given for each day, however please check our
Attributions section for efforts beyond the lab work.


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