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Aims

The sensitivity of the system is to be determined through this experiment. To do this, we induce the system with known concentrations of AHL. We then record the change in GFP, such that we can calculate the rate of GFP production relative to concentration of AHL in solution.

• To determine the rate of change of protein production relative to concentration of AHL
• To determine the response time relative to concentration of AHL
• To determine the steady state response of protein relative to concentration of AHL

Materials and Methods

AHL Dilutions

8 concentrations of AHL were tested - 0, 5, 10, 15, 20, 50, 100, 1000 nM. These concentrations of AHL were produced by serial dilutions. Indepth protocols of these dilutions are documented at AHL dilutions page.

Preparing Cell Extract

The following mixture was made

• 20µl Premix Solution
• 15µl S30 Cell Extract
• 2.5µl Amino Acid Mixture 1
• 2.5µl Amino Acid Mixture 2
• 10.2µl DNA (4 µg)
• 9.8µl Nuclease-Free Water

Further details on protocols... ... (page pending completion)

Testing and Results

Discussion and Validation