Team:BIOTEC Dresden/Protocols:Cocultivation assay

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<h2>Set-up</h2>

<h3>Tecan plate reader</h3>
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<li>Start the machine <span class="markup time">20 minutes</span> in advance and heat the chamber to <span class="markup temp">37°C</span>. Put in the plate and start the measurement with the following setting:</li>
<li>Measure OD at 612 nm</li>
<li>Measure fluorescence with an excitation wavelength of 485nm and an emission wavelength of 535nm for both GFP and YFP</li>
<li>Shake</li>
<li>Repeat measurements every <span class="markup time">5min for 3 hours</span></li>
</ul>

<a rel="lightbox"><img class="border" style="width:300px; margin-left:20%" src="https://static.igem.org/mediawiki/2010/1/17/BiotecDresden_Pipetting_schema_cocultivation_l.png"></a>
<h3>Data processing</h3>
<ul>
<li>Normalize the data by dividing all fluorescence data by the corresponding optical density</li>
<li>Subtract the obtained value of the reference column from the calculated relative fluorescence of the wells containing HHL</li>
<li>Plot the fluorescence data over the time</li>
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[[Category:BIOTEC Dresden/Protocol|Cocultivation assay]]
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