Hrp Characterisation: Testing
- The motvation behind the characterisations carried out is to create well characterised parts and devices, and to characterise in terms of; GFP molecules synthesized cfu -1 sec-1 .
General tasks to be completed for each of our constructs:
- Amplifying the biobricks
- Cloning of the constructs
- Growing Cultures
- Measuring Inputs - GFP downstream of Promoters
- Measuring Outputs- GFP downstream of Devices
1. Amplifying the biobricks
2. Cloning of the Constructs
4. Growing Cultures
- The fluorometer used in these experiments is 'The Twinkle'.
- This machine is a plate reader fluorometer that can measure 96 well plates.
- Using this machine it is hoped that absorbance at 600nm and the fluorescence emission of acGFP can be measured at set time intervals.
- The machine is being installed 3rd of August 10.30am.
- The general principle is to measure the fluorescence into the device and out of the device, this allows characteristics such as transfer function to be calculated.
- To enable us to measure inputs and output, two separate experiments will be carried out, one for the inputs fluorescence and one for the output of fluorescence.
- A Hrp Protocol can be found here. This is a general protocol and can be applied to the various devices and parts to be characterised.
- Two specific fluorescence measurements will be made for each device:
- Input -The input of fluorescence will be measured at varying inducer concentrations, to give the value of GFP molecules synthesized cfu -1 sec -1 into our device at specific inducer concentrations.
- Output -The output of fluorescence will be measured at varying inducer concentrations and at various time intervals. The inducer concentration will have a known GFP molecules synthesized cfu -1 sec -1 input and so we will be measuring the time response and level of output of our device.
- The two measurements that will be made for the various devices are fluorescence emission and absorbance.
- The aim of the characterisation is to use these two measurements in order to create a more generic unit to allow for modular design and easier repetition of experiments. The unit that has been chosen to be used is GFP molecules synthesised cfu -1 sec-1 .
- In order to convert our raw data into these units, two sets of calibration curves are needed.
- Here is a link to an explanation of the Calibration Curves