Team:BIOTEC Dresden/Protocols:Cocultivation assay
From 2010.igem.org
(Difference between revisions)
Line 13: | Line 13: | ||
<li>Repeat measurements every <span class="markup time">5min for 3 hours</span></li> | <li>Repeat measurements every <span class="markup time">5min for 3 hours</span></li> | ||
</ul> | </ul> | ||
+ | |||
+ | <a rel="lightbox" class="border" style="width:300px; margin-left:20%"><img 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> |
Revision as of 02:49, 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