Imperial/Wet Lab/Results/CBD2.3

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== Results ==
== Results ==
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[[Image:IC 2007 CBD steps.PNG|600px|center|thumb|Fig 1.1:Molecules of GFPmut3b synthesised on average over time for two samples at constant temperatures of 4&degC and 20&deg;C and two with a temperature step (4-20&deg;C and 20-4&deg;C)]]   
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| width="600px"|<br>[[Image:IC 2007 CBD steps.PNG|600px|Fig 1.1:Average fluorescence over time for two samples at constant temperatures of 4&degC and 20&deg;C and two with a temperature step (4-20&deg;C and 20-4&deg;C)]]   
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'''Raw Data:'''
'''Raw Data:'''
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*[[Media:IC 2007 CBD steps.xls|Raw Data excel file]]
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*[[Media:CBD steps.xls|Raw Data excel file]]
== Discussion ==
== Discussion ==
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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.  
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The 'step' samples were moved from their original temperature to the other temperature 180 mins 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.  
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Both samples at 4&deg;C have similar fluorescence until 180mins. After the sample is moved to 20&deg;C, the fluorescnce produced increases to a very high value.
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Both samples at 4&deg;C have similar number of molecules of GFPmut3b produced until 180 mins. After one of the samples is moved to 20&deg;C, the GFPmut3b molecules produced in that sample increased to a very high value.
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Moving the sample at 20&deg;C to 4&deg; 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.  
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Moving the sample at 20&deg;C to 4&deg; decreased the amount of fluorescent protein synthesized by a small amount. This is what was expected as the pTet-GFPmut3b construct showed linear relationship with temperature in the earlier experiments.  
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== Conclusion ==
== Conclusion ==
It can be concluded from the results that the pTet-GFPmut3b construct is sensitive to a step increase and step decrease in the temperature.
It can be concluded from the results that the pTet-GFPmut3b construct is sensitive to a step increase and step decrease in the temperature.

Latest revision as of 02:53, 27 October 2007




In vitro Testing of pTet-GFPmut3b Construct for constant and step temperatures

Aims

To determine how the system ([http://partsregistry.org/Part:BBa_I13522 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

Results

Fig 1.1:Molecules of GFPmut3b synthesised on average over time for two samples at constant temperatures of 4&degC and 20°C and two with a temperature step (4-20°C and 20-4°C)


Controls:

  • Negative control - empty vector (pTet promoter) with the cell extract


Constants:

  • Temperature - 4°C and 20°C
  • Total Volume - 60µl


Raw Data:

Discussion

The 'step' samples were moved from their original temperature to the other temperature 180 mins 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 number of molecules of GFPmut3b produced until 180 mins. After one of the samples is moved to 20°C, the GFPmut3b molecules produced in that sample increased to a very high value.

Moving the sample at 20°C to 4° decreased the amount of fluorescent protein synthesized by a small amount. This is what was expected as the pTet-GFPmut3b construct showed linear relationship with temperature in the earlier experiments.

Conclusion

It can be concluded from the results that the pTet-GFPmut3b construct is sensitive to a step increase and step decrease in the temperature.