Difference between revisions of "Part:BBa K1073024:Experience"

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When expressed the chromoprotein amilGFP exhibits a strong yellow color. In order to avoid absorption of the chomoproteins during measurement with the photometer the absorption and emission spectra were measured and an optimal wavelength for spectral optical density measurement was determined according to these spectra.
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When expressed the chromoprotein amilGFP exhibits a strong yellow color. In order to avoid absorption by the chomoproteins during the measurement of the OD of bacterial cultures, the absorption and emission spectra were measured and an optimal wavelength for spectral optical density measurement was determined to be at 520nm.
  
 
[[File:BSAbsorption-amilGFP.jpg |400px]]
 
[[File:BSAbsorption-amilGFP.jpg |400px]]

Revision as of 11:48, 28 September 2013


This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K1073024

When amilGFP is expressed, the colonies on plates as well as the liquid culture become yellow.

User Reviews

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iGEM Team Braunschweig 2013

When expressed the chromoprotein amilGFP exhibits a strong yellow color. In order to avoid absorption by the chomoproteins during the measurement of the OD of bacterial cultures, the absorption and emission spectra were measured and an optimal wavelength for spectral optical density measurement was determined to be at 520nm.

BSAbsorption-amilGFP.jpg

iGEM Team Braunschweig 2013: Absorption spectrum of chromoprotein amilGFP in the wavelength range 400-800 nm.

Absorption maximum of amilGFP: 504 nm

BS2013Emission amilGFP.jpg

iGEM Team Braunschweig 2013: Emission spectrum of chromoprotein amilGFP. This chromoprotein does show significant emission and can be detected by its fluorescence as well as by naked eye.

Emission maximum of amilGFP: 515,8 nm


According to these results the optical density for growth curves of cells expressing this protein should be measured at 520 nm with respect to the chromoproteins eforRed (BBa_K1073022) and aeBlue (BBa_K1073020) to avoid absorption by the proteins. Comparing all absorption spectra they all show an absorption minimum at 520 nm.

Although this device is working perfectly the way it is expected to the iGEM Team Braunschweig 2013 did not use it because the yellow color is not as easiely distinguishable by naked eye from the color of the media as the other chromoproteins are.

Compraring chromoproteins.jpg

iGEM Team Braunschweig 2013: Supernatant after cell disruption containing chromoproteins.


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