IGEM:IMPERIAL/2007/Experimental Design/Phase2/Results 1.1: Difference between revisions

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==Experiment==
= Fluorescence Dependence on Media Volume=
'''Aims'''
*To determine if construct expresses in vitro at temperatures of: 4oC, 15oC, 25oC, 30oC, 37oC, 50oC
*To determine how long the system can last (when GFP is no longer produced) at each temperature.
*To determine the maximum rate of GFP produced at each temperature range by taking fluoresence reading over a period of time ( time frame to be determined by Friday 17th August)
'''Status'''<br>
???<br>
'''Equipments'''
*Fluorometer + PC
*Fridge at 4oC
*Water bath in cold room at 15oC
*30oC water bath
*37oC shaking incubator
*50oC heating block
*6 Fluorometer plates (black)
*6 Sealing plate mats
*Gilson pipettes 1000, 200 and 20
*Eppendorf Tubes
*Plate Centrifuge
*Stopwatch


'''Steps'''
__NOTOC__
==Aims==
To investigate the fluorescence dependence of varying media volumes to no. of moles of GFP. This is to determine the need for the maintenance of a constant volume throughout subsequent experiments, as with making GFP dilutions for the calibration curve.
 
 
==Materials and Methods==
Refer to protocols page.
 
==Results==
[[Image:IC2007 Experimental Design GFP Dilution in water.PNG|thumb|left|800px| Fig.1: Fluorescence of varying media volume]]
<br>
 
Interestingly, the addition of more media (water) increased the fluorescence readings of the samples to a significant extent (Fig.1).
 
 
==Discussion==
Fig.1 suggests that fluorescence does depend on the wolume of media to which the substrate is diluted in, albeit in a proportionate way. While one would expect the opposite to occur, the results suggest that volume of media, and hence the maintenance of the same volume through subsequent experiments, is vital for an accurate reading. Indeed this also implicitly implies the importance of minimizing evaporation in our media.
 
 
==Conclusion==
* Constant volume must be maintained for all experiments.
* Evaporation (reducing media volume) may play a huge role in experimental readings.

Latest revision as of 04:04, 16 October 2007

Fluorescence Dependence on Media Volume

Aims

To investigate the fluorescence dependence of varying media volumes to no. of moles of GFP. This is to determine the need for the maintenance of a constant volume throughout subsequent experiments, as with making GFP dilutions for the calibration curve.


Materials and Methods

Refer to protocols page.

Results

Fig.1: Fluorescence of varying media volume


Interestingly, the addition of more media (water) increased the fluorescence readings of the samples to a significant extent (Fig.1).


Discussion

Fig.1 suggests that fluorescence does depend on the wolume of media to which the substrate is diluted in, albeit in a proportionate way. While one would expect the opposite to occur, the results suggest that volume of media, and hence the maintenance of the same volume through subsequent experiments, is vital for an accurate reading. Indeed this also implicitly implies the importance of minimizing evaporation in our media.


Conclusion

  • Constant volume must be maintained for all experiments.
  • Evaporation (reducing media volume) may play a huge role in experimental readings.