In vitro Testing of pTet-GFPmut3b Construct for constant and step temperatures
To determine how the system (pTet-GFPmut3b) responds to a step increase or a step decrease in temperature, compared to a constant temperture, in vitro.
Materials and Methods
Link to the Protocols page
- Negative control - empty vector (pTet promoter) with the cell extract
- Temperature - 4°C and 20°C
- Total Volume - 60µl
The 'step' samples were moved from their original temperature to the other 180mins after the start of the experiment. As it can be seen from the results above, the temperature step has a very rapid affect on the fluorescence of the DNA construct.
Both samples at 4°C have similar fluorescence until 180mins. After the sample is moved to 20°C, the fluorescnce produced increases to a very high value.
Moving the sample at 20°C to 4° seems to have increased the fluorescence by a little amount. This is not what was expected as the pTet-GFPmut3b construct showed linear relationship with temperature earlier.
It can be concluded from the results that the pTet-GFPmut3b construct is sensitive to a step increase and step decrease in the temperature.