IGEM:IMPERIAL/2007/Experimental Design/Phase1/Results 2.2: Difference between revisions

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====Testing at 25 <sup>o</sup>C ====
====Testing at 25 <sup>o</sup>C ====
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Today we tested pLux in vitro using 3 different combinations of cell extract:
<br>
# pLux with Commercial(CM) S30 cell extract and CM Premix
# pLux with Homemade(HM) S30 cell extract and HM Premix
# pLux with HM S30 cell extract and CM Premix
<br>
The idea behind the first combination is to see if pLux works in vitro since we already know our commercial extract is working with pTet. If pLux does work with the commercial extract but proves to be reduntant in the second combination, which utilises only homemade cell extract & premix, then we know that our homemade extract inefficient. Lastly, the third combination is to test whether our homemade cell extract will work better with the commercial premix rather than the homemade one.
The experiment was carried out over a period of nearly 6:30 hours, taking measuremets every half hour. Each combination had 3 repeats and 1 control. The average value of fluorescence for each combination is on the graph below:
<br>
[[ Image:IC2007 Experimental Design PLux vitro 6hours.PNG|thumb|left|500px| Results of in vitro testing of pLux over a 6 hour period at 25<sup>o</sup>C ]]
<br>
'''The graph on the right dispays the following.'''
<div style="color:Green">Fluorescence of the diluted GFP solution (+ve control).</div>
<div style="color:#F88017">The average fluorescence of the pLux samples(3) in CM cell extract + CM premix</div>
<div style="color:lightBlue">The average fluorescence of the pLux samples(3) in HM cell extract + HM premix</div>
<div style="color:Black">The average fluorescence of the pLux samples(3) in HM cell extract + CM premix</div>
<div style="color:#C45AEC">The fluoresence of a solution containing only S30 cell extract and nuclease free water (-ve control) .</div>
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Revision as of 03:25, 6 September 2007


Testing at 25 oC

Today we tested pLux in vitro using 3 different combinations of cell extract:

  1. pLux with Commercial(CM) S30 cell extract and CM Premix
  2. pLux with Homemade(HM) S30 cell extract and HM Premix
  3. pLux with HM S30 cell extract and CM Premix


The idea behind the first combination is to see if pLux works in vitro since we already know our commercial extract is working with pTet. If pLux does work with the commercial extract but proves to be reduntant in the second combination, which utilises only homemade cell extract & premix, then we know that our homemade extract inefficient. Lastly, the third combination is to test whether our homemade cell extract will work better with the commercial premix rather than the homemade one.

The experiment was carried out over a period of nearly 6:30 hours, taking measuremets every half hour. Each combination had 3 repeats and 1 control. The average value of fluorescence for each combination is on the graph below:

Results of in vitro testing of pLux over a 6 hour period at 25oC


The graph on the right dispays the following.

Fluorescence of the diluted GFP solution (+ve control).
The average fluorescence of the pLux samples(3) in CM cell extract + CM premix
The average fluorescence of the pLux samples(3) in HM cell extract + HM premix
The average fluorescence of the pLux samples(3) in HM cell extract + CM premix
The fluoresence of a solution containing only S30 cell extract and nuclease free water (-ve control) .



Experiment 1

Experiment 2

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