Degradation Curve: Fluroescence of GFPmut3b as a function of time
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
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.