Imperial/Cell by Date/Testing
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= Cell by Date: Testing = | = Cell by Date: Testing = | ||
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==Summary== | ==Summary== | ||
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| + | Having determined that our system worked at a basic level in our [https://2007.igem.org/Imperial/Cell_by_Date/Implementation implementation phase] we set out to ascertain a clearer picture of the performance of our system to ultimately reveal whether our design had realised our specifications. | ||
==Results== | ==Results== | ||
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'''Other practical considerations:''' <br> | '''Other practical considerations:''' <br> | ||
*Unfortunetly we have not been able to use a visible reporter and so have no idea whether the construct can in fact produce a visible signal. <br> | *Unfortunetly we have not been able to use a visible reporter and so have no idea whether the construct can in fact produce a visible signal. <br> | ||
| - | *In terms of lifespan we have had major problems with evaporation meaning that the lifespan of our system is limited to a few days. <br> | + | *In terms of lifespan we have had major problems with evaporation meaning that the lifespan of our system is limited to a few days as discussed in our packaging experiment below. <br> |
===<font color=blue>Other Experiments</font>=== | ===<font color=blue>Other Experiments</font>=== | ||
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2. Packaging | 2. Packaging | ||
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Revision as of 00:59, 27 October 2007
Cell by Date: Testing
Summary
Key results of our experiments are :
1.Operating Range : Our System Operates between 8-37 Degrees
2.Activation Energy : The activation energy of our system is 1.5kJ/mol
3.DNA Concentration : The optimum DNA concentration for our system is 2-4ug DNA
Aims
Having determined that our system worked at a basic level in our implementation phase we set out to ascertain a clearer picture of the performance of our system to ultimately reveal whether our design had realised our specifications.
Results
Investigating properties of system under isothermal scenarios
To estimate the properties of our system we looked at the evolution of the Fluorescence with time for experiments where temperature is carefully kept constant. (Figure 1).
General Behaviour:
- In all experiment we can observe a linear growth of fluorescence corresponding to a steady rate of protein production.
- This matches the claims of the manufacturer of the cell-free extract that the extract is optimised so that protein degradation is negligible.
Operating Range:
- Our system seems to 'turn off' at around 4 degrees C.
- At 8 degrees with have minimal expression which then increases to a substantial expression at 37 degrees.
Synthesis rate vs Temperature
- Over the range of temperature considered here the rate increases with temperature.
- To investigate the relation between synthesis rate and temperature we extracted the rate of synthesis of GFP for each experiment and plotted it against 1/T (Figure 2).
- A strong linear correlation in the log plot supports an arrhenius type dependence on temperature with an activation energy of 1.5kJ/mol.
Investigating properties of system under isothermal scenarios
Other practical considerations:
- Unfortunetly we have not been able to use a visible reporter and so have no idea whether the construct can in fact produce a visible signal.
- In terms of lifespan we have had major problems with evaporation meaning that the lifespan of our system is limited to a few days as discussed in our packaging experiment below.
Other Experiments
1. DNA Concentration
2. Packaging
