IGEM:Imperial/2007/Wet Lab/results/Res1.4: Difference between revisions
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To construct a degradation curve of GFPmut3b in the in vitro chassis. This will verify whether the degradation of the system would be a limiting factor to the expression of the system. | To construct a degradation curve of GFPmut3b in the in vitro chassis. This will verify whether the degradation of the system would be a limiting factor to the expression of the system. | ||
==Materials and methods== | |||
Link to the [http://parts.mit.edu/igem07/index.php/Imperial/Wet_Lab/Protocols/Prot1.4 Protocol] | Link to the [http://parts.mit.edu/igem07/index.php/Imperial/Wet_Lab/Protocols/Prot1.4 Protocol] | ||
==Results== | ==Results== | ||
[[Image:IC 2007 Degradation.PNG|left|thumb|500px|Figure 1.1 ]] | [[Image:IC 2007 Degradation.PNG|left|thumb|500px|Figure 1.1 Fluorescence vs time]] | ||
[[Image:IC 2007 Degradation log.png|right|thumb|500px|Figure 1.2 Semi-log plot of fluorescence vs time]] | |||
<br clear="all"> | |||
==Conclusion== | |||
The degradation constant was calculated from the semi-log plot. The average gradients of all the semi-log plots was -0.0003501. This is equivalent to a decay-constant of 0.0003501. Using the equation ln2/k = t<sub>1/2</sub>, the half-life of GFP was calculated to 1980 minutes, or approximately 33 hours. This agrees with the literature value of ~24 hours. So for our experimental timescale of 6 hours, we would only observe 12% of our GFP to be degraded. This supports our assumption that the degradation of GFP is very small within the timescale of our experiments. | |||
'''Raw Data''' | '''Raw Data''' | ||
*[[ | *[[Media:IC 2007 Degradation curve 25oC 04-10.xls| Raw Data excel file ]] | ||
Latest revision as of 08:19, 24 October 2007
Degradation Curve: Fluroescence of GFPmut3b as a function of time
Aims
To construct a degradation curve of GFPmut3b in the in vitro chassis. This will verify whether the degradation of the system would be a limiting factor to the expression of the system.
Materials and methods
Link to the Protocol
Results
Conclusion
The degradation constant was calculated from the semi-log plot. The average gradients of all the semi-log plots was -0.0003501. This is equivalent to a decay-constant of 0.0003501. Using the equation ln2/k = t1/2, the half-life of GFP was calculated to 1980 minutes, or approximately 33 hours. This agrees with the literature value of ~24 hours. So for our experimental timescale of 6 hours, we would only observe 12% of our GFP to be degraded. This supports our assumption that the degradation of GFP is very small within the timescale of our experiments.
Raw Data
