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Team:BIOTEC Dresden/Protocols:Cocultivation assay
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<h3>Tecan plate reader</h3> | <h3>Tecan plate reader</h3> | ||
<ul> | <ul> | ||
| - | <li>Start the machine 20 minutes in advance and heat the chamber to | + | <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 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>Measure fluorescence with an excitation wavelength of 485nm and an emission wavelength of 535nm for both GFP and YFP</li> | ||
<li>Shake</li> | <li>Shake</li> | ||
| - | <li>Repeat measurements every 5min for 3 hours</li> | + | <li>Repeat measurements every <span class="markup time">5min for 3 hours</span></li> |
</ul> | </ul> | ||
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| + | <a rel="lightbox"><img class="border" style="width:500px; margin-left:20%" src="https://static.igem.org/mediawiki/2010/1/17/BiotecDresden_Pipetting_schema_cocultivation_l.png"></a> | ||
<h3>Data processing</h3> | <h3>Data processing</h3> | ||
<ul> | <ul> | ||
Latest revision as of 02:51, 28 October 2010
Set-up
Tecan plate reader
- Start the machine 20 minutes in advance and heat the chamber to 37°C. Put in the plate and start the measurement with the following setting:
- Measure OD at 612 nm
- Measure fluorescence with an excitation wavelength of 485nm and an emission wavelength of 535nm for both GFP and YFP
- Shake
- Repeat measurements every 5min for 3 hours
Data processing
- Normalize the data by dividing all fluorescence data by the corresponding optical density
- Subtract the obtained value of the reference column from the calculated relative fluorescence of the wells containing HHL
- Plot the fluorescence data over the time
