Part:BBa_K1586000:Experience

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Characterisation

Experimental:
In order to characterise that this part works as expected, fluorescence intensity was measured for the expression of K1586000 in a cell free system in the presence of different amounts of trigger RNA, where the plasmid encoding for the toehold was kept constant at 0.5 pmols. Below is a graph showing a positive correlation between amount of trigger RNA (in log10(nanograms) with 0ng normalised to 0) and GFP fluorescence intensity.

In silico:
As well as generating experimental data in the lab, in silico data was also generated through the software package NUPACK using free energies. For a full explanation of how this works, please visit http://2015.igem.org/Team:Exeter/Modeling.

The equilibrium concentrations of the different components in our system were determined, along with free energy data for the different structures; the unbound trigger, the unbound toehold and the complex of the two.




After obtaining this, the data was converted to an approximate normal distribution and after normalising these within the range of values, we could use the distribution as a probability distribution. Shown below are these obtained probability distributions based on the free energy data.

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