Team:Braunschweig/Notebook

From 2013.igem.org

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Pre-cultures of chromoprotein constructs were mixed in one main culture in order to see how they behave during cultivation over several hours. OD<sub>520</sub> of pre-cultures was measured in order to inoculate the main culture with  33% of each strain to a final OD<sub>520</sub>=0.3. For the main culture, 25 ml 2xYT containing chloramphenicol were inoculated with the three different strains and grown at 37°C and 250 rpm in a non-baffled flask. Samples from the culture were taken at several time points, diluted and plated on 2xYT agar-plates containing chloramphenicol. Agar plates were incubated at 37°C over night.<br>
Pre-cultures of chromoprotein constructs were mixed in one main culture in order to see how they behave during cultivation over several hours. OD<sub>520</sub> of pre-cultures was measured in order to inoculate the main culture with  33% of each strain to a final OD<sub>520</sub>=0.3. For the main culture, 25 ml 2xYT containing chloramphenicol were inoculated with the three different strains and grown at 37°C and 250 rpm in a non-baffled flask. Samples from the culture were taken at several time points, diluted and plated on 2xYT agar-plates containing chloramphenicol. Agar plates were incubated at 37°C over night.<br>
Today we also observed our cells containing the chromoprotein expression cassette under the fluorescence microscope.
Today we also observed our cells containing the chromoprotein expression cassette under the fluorescence microscope.
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<img alt="microscope" src="https://static.igem.org/mediawiki/2013/0/09/Braunschweig_Lab_Week13_microscope.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/2013/0/09/Braunschweig_Lab_Week13_microscope.jpg" height="220px" align="left" vspace="0" hspace="10" style="margin-right:5px"/>
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During the day we noticed that the 2xYT medium used for the pre-cultures yesterday showed contamination. Thus, there is a high possibility  of contamination in the pre-culture as well as in the main culture. Therefore we decided to repeat the experiment. New pre-cultures of the three chromoprotein expression cassettes where inoculated in 50 ml 2xYT containing chloramphenicol directly from -80°C.<br>
During the day we noticed that the 2xYT medium used for the pre-cultures yesterday showed contamination. Thus, there is a high possibility  of contamination in the pre-culture as well as in the main culture. Therefore we decided to repeat the experiment. New pre-cultures of the three chromoprotein expression cassettes where inoculated in 50 ml 2xYT containing chloramphenicol directly from -80°C.<br>
In order to have our constructs available in different <i>E. coli</i> strains for the fluorescence microscopy experiments, the eforRed expression cassette was transformed into <i>E. coli</i> Top10F' by electroporation. Transformed cells were plated on 2xYT agar containing chloramphenicol and incubated over night at 37°C.<br>
In order to have our constructs available in different <i>E. coli</i> strains for the fluorescence microscopy experiments, the eforRed expression cassette was transformed into <i>E. coli</i> Top10F' by electroporation. Transformed cells were plated on 2xYT agar containing chloramphenicol and incubated over night at 37°C.<br>
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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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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.</p>
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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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<img alt="continuous 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"/></p>
<p><img alt="grey line" src="https://static.igem.org/mediawiki/2013/4/4c/Braunschweig_grey_line.png" width="850" height="1" vspace="20"/></p>
<p><img alt="grey line" src="https://static.igem.org/mediawiki/2013/4/4c/Braunschweig_grey_line.png" width="850" height="1" vspace="20"/></p>

Revision as of 23:18, 4 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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