Difference between revisions of "Team:Heidelberg/Collaborations"

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            <h1>1. Interlab Study</h1>
  
Mutagenesis plasmids are crucial to enable rapid mutation that makes continuous evolution possible in short time scales. The mutagenesis plasmids we used have a \(P_{BAD}\) promotor that is arabinose inducible but suppressed by glucose<x-ref>RN159</x-ref>. Consequently controlling the glucose concentration is important in order to have a strong induction of the mutagenesis plasmids that results in a high mutation rate, which leads to a larger covered sequence space. This model uses ordinaray differential equations (ODE) to model both the glucose and the <i>E. coli</i> concentration, assuming both are independend of each other. This is plausible because <a href="https://2017.igem.org/Team:Heidelberg/Experiments#medium">the medium used in the experiments</a> contained other carbon sources than glucose. The glucose consumption rate per <i>E. coli</i> is assumed to be independent of the glucose concentration.
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            As a first step to contribute to this year's iGEM competition we decided to participate in iGEM's fourth International Interlaboratory Study along with many other teams from around the world. This study is organized by iGEM's measurement committee in an effort to establish standardized, reliable and repeatable measurement tools for the iGEM community and the synthetic biology community as a whole. This year's iGEM InterLab study is about establishing a standardized protocol for the measurement of GFP using a plate reader. To start things off we needed a plate reader that is qualified to measure GFP fluorescence. Namely, Tecan Infinite M200 Pro plate reader. Additionally, we needed competent <i>E.coli</i> DH5&#945;. These were prepared from glycerol stocks. Together with the material iGEM had provided we were ready for work. Throughout our experiments, we tested 8 plasmids (2 controls and 6 test devices) by measuring the OD<sub>600</sub> and the fluorescence of the cells carrying the constructs.
 
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            The workflow can be separated into four segments. The first segment is the transformation of all plasmids into competent  DH5&#945; cells.
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The arabinose concentration is easier to tune since higher arabinose concentrations induce the mutagenesis plasmids stronger and have no known negative side effects. This shifts the focus of the induction modeling to the glucose concentration.
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Revision as of 21:52, 29 October 2017


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1. Interlab Study

As a first step to contribute to this year's iGEM competition we decided to participate in iGEM's fourth International Interlaboratory Study along with many other teams from around the world. This study is organized by iGEM's measurement committee in an effort to establish standardized, reliable and repeatable measurement tools for the iGEM community and the synthetic biology community as a whole. This year's iGEM InterLab study is about establishing a standardized protocol for the measurement of GFP using a plate reader. To start things off we needed a plate reader that is qualified to measure GFP fluorescence. Namely, Tecan Infinite M200 Pro plate reader. Additionally, we needed competent E.coli DH5α. These were prepared from glycerol stocks. Together with the material iGEM had provided we were ready for work. Throughout our experiments, we tested 8 plasmids (2 controls and 6 test devices) by measuring the OD600 and the fluorescence of the cells carrying the constructs. The workflow can be separated into four segments. The first segment is the transformation of all plasmids into competent DH5α cells.