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Team:DTU-Denmark/Achievements/Modelling
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When the fluorophore DFHBI is bound by Spinach, it fluoresces much more intensely than in its unbound form. The fluorescence of a sample can be assumed to increase linearly with the concentration of Spinach-DFHBI complex:<br> | When the fluorophore DFHBI is bound by Spinach, it fluoresces much more intensely than in its unbound form. The fluorescence of a sample can be assumed to increase linearly with the concentration of Spinach-DFHBI complex:<br> | ||
<img src="https://static.igem.org/mediawiki/2014/1/15/DTU-Denmark_modelling_equation1.png" class="modelling_equation" /> | <img src="https://static.igem.org/mediawiki/2014/1/15/DTU-Denmark_modelling_equation1.png" class="modelling_equation" /> | ||
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By adding known concentrations of DFHBI to an excess amount of Spinach RNA, it is possible to make a standard series describing the correlation between fluorescence and DFHBI concentration, and estimate the parameters <i>a</i> and <i>b</i>, if it is assumed that DFHBI is bound completely by the excess Spinach, i.e.:<br> | By adding known concentrations of DFHBI to an excess amount of Spinach RNA, it is possible to make a standard series describing the correlation between fluorescence and DFHBI concentration, and estimate the parameters <i>a</i> and <i>b</i>, if it is assumed that DFHBI is bound completely by the excess Spinach, i.e.:<br> | ||
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Revision as of 21:29, 16 October 2014
- Home
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At the Giant Jamboree, we received a gold medal and an Interlab Study Award. We also won the Overgrad Best Parts Collection.
The main objective of the experimental part of our project was to develop a method for determining promoter activities by measuring fluorescence from the Spinach RNA-fluorophore complex.
Fluorescence Signal is Linearly Dependent on the Concentration of Spinach-DFHBI complex
When the fluorophore DFHBI is bound by Spinach, it fluoresces much more intensely than in its unbound form. The fluorescence of a sample can be assumed to increase linearly with the concentration of Spinach-DFHBI complex:
where the intercept b is the background fluorescence without any Spinach-DFHBI complex, and the slope a is the increase in fluorescence for each concentration unit of Spinach-DFHBI complex.
By adding known concentrations of DFHBI to an excess amount of Spinach RNA, it is possible to make a standard series describing the correlation between fluorescence and DFHBI concentration, and estimate the parameters a and b, if it is assumed that DFHBI is bound completely by the excess Spinach, i.e.:
This can be used to calculate the concentration of the Spinach-DFHBI complex given a fluorescence measurement:
[EQUATION_3]
If the fluorescence of a Spinach-expressing culture is measured, the standard series can thus be used to calculate the concentration of Spinach-DFHBI complex in the culture.
Calculating Total Spinach Concentration in the Culture
If the fluorescence is measured in the culture with a large excess of DFHBI it can be assumed that the concentration of Spinach-DFHBI is equal to the concentration of correctly folded Spinach, i.e.:
Technical University of Denmark
Department of Systems Biology
Søltofts Plads 221
2800 Kgs. Lyngby
Denmark
+45 45 25 25 25
dtuigem14@gmail.com .
+45 45 25 25 25
dtuigem14@gmail.com . 
