Imperial/Wet Lab/Results/CBD2.3
From 2007.igem.org
m (→Results) |
(→Discussion) |
||
| (13 intermediate revisions not shown) | |||
| Line 1: | Line 1: | ||
| - | |||
| + | |||
| + | {{Template: IC07navmenu}} | ||
| + | <br clear="all"> | ||
__NOTOC__ | __NOTOC__ | ||
| + | |||
| + | = ''In vitro'' Testing of pTet-GFPmut3b Construct for constant and step temperatures= | ||
| + | |||
==Aims== | ==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''. | 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''. | ||
| Line 9: | Line 14: | ||
== Results == | == Results == | ||
| - | |||
| - | |||
| - | |||
| + | [[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°C and 20°C and two with a temperature step (4-20°C and 20-4°C)]] | ||
<br clear="all"> | <br clear="all"> | ||
| Line 23: | Line 26: | ||
<br clear="all"> | <br clear="all"> | ||
'''Raw Data:''' | '''Raw Data:''' | ||
| - | *[[Media: | + | *[[Media:CBD steps.xls|Raw Data excel file]] |
== Discussion == | == Discussion == | ||
| - | The 'step' samples were moved from their original temperature to the other | + | 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 | + | 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° | + | 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. |
<br clear="all"> | <br clear="all"> | ||
== 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
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.
