"
Team:BIOTEC Dresden/Protocols:Cocultivation assay
From 2010.igem.org
(Difference between revisions)
(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...) |
|||
| (4 intermediate revisions not shown) | |||
| Line 1: | Line 1: | ||
{{Biotec_Dresden/Header}} | {{Biotec_Dresden/Header}} | ||
<html> | <html> | ||
| + | <head> | ||
| + | |||
| + | <script type="text/javascript" src="http://endo.bio.tu-dresden.de/igem/scripthost/fancybox/jquery.easing-1.3.pack.js"></script> | ||
| + | <script type="text/javascript" src="http://endo.bio.tu-dresden.de/igem/scripthost/fancybox/jquery.fancybox-1.3.2.pack.js"></script> | ||
| + | <link rel="stylesheet" href="http://endo.bio.tu-dresden.de/igem/scripthost/fancybox/jquery.fancybox-1.3.2.css" type="text/css" media="screen" /> | ||
| + | <script type="text/javascript"> | ||
| + | $(document).ready(function() { | ||
| + | $("a[rel='lightbox']").fancybox({ | ||
| + | 'transitionIn' : 'elastic', | ||
| + | 'transitionOut' : 'elastic', | ||
| + | 'speedIn' : 600, | ||
| + | 'speedOut' : 200, | ||
| + | 'overlayShow' : false, | ||
| + | 'titleShow': false | ||
| + | }); | ||
| + | }); | ||
| + | </script> | ||
| + | </head> | ||
<body> | <body> | ||
<div id="content_prim"> | <div id="content_prim"> | ||
| Line 6: | Line 24: | ||
<h3>Tecan plate reader</h3> | <h3>Tecan plate reader</h3> | ||
| - | + | <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> | ||
| + | <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> | |
<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> | ||
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
