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Team:BIOTEC Dresden/Protocols:Cocultivation assay
From 2010.igem.org
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(New page: {{Biotec_Dresden/Header}} <html> <body> <div id="content_prim"> <h2>Set-up</h2> <h3>Tecan plate reader</h3> <li>Start the machine 20 minutes in advance and heat the chamber to 37C. Pu...) |
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<h3>Tecan plate reader</h3> | <h3>Tecan plate reader</h3> | ||
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<li>Start the machine 20 minutes in advance and heat the chamber to 37C. Put in the plate and start the measurement with the following setting:</li> | <li>Start the machine 20 minutes in advance and heat the chamber to 37C. 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 5min for 3 hours</li> | ||
| - | + | </ul> | |
<h3>Data processing</h3> | <h3>Data processing</h3> | ||
| - | + | <ul> | |
<li>Normalize the data by dividing all fluorescence data by the corresponding optical density</li> | <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>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> | <li>Plot the fluorescence data over the time</li> | ||
| + | </ul> | ||
</div> | </div> | ||
</body> | </body> | ||
Revision as of 17:03, 26 October 2010
Set-up
Tecan plate reader
- Start the machine 20 minutes in advance and heat the chamber to 37C. 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
