IGEM:IMPERIAL/2007/Experimental Design/Phase1/Results 3.1: Difference between revisions
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To test which is the optimum time for us to use in our experiments we followed the protocol found in the previous page. For each time point examined, different samples of the same stock solution where measured repeatedly(4 times) at different windows(counting time). From there we calculated, for each window, the percentage(%) variability between the repeated measurements(4) of the same sample. Ideally, we since the same sample is measured repeatedly, we expect the variability between the repeats to be zero(0). This however does not take into consideration the randomness inherent within fluorescence. The results are as follows: | To test which is the optimum time for us to use in our experiments we followed the protocol found in the previous page. For each time point examined, different samples of the same stock solution where measured repeatedly(4 times) at different windows(counting time). From there we calculated, for each window, the percentage(%) variability between the repeated measurements(4) of the same sample. Ideally, we since the same sample is measured repeatedly, we expect the variability between the repeats to be zero(0). This however does not take into consideration the randomness inherent within fluorescence. The results are as follows: | ||
[[ Image:IC2007 Experimental Design Phase1 Protocol31Window-experiment.PNG|thumb| | [[ Image:IC2007 Experimental Design Phase1 Protocol31Window-experiment.PNG|thumb|800px|left]] | ||
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The most optimum window length to be used is found to be the 0.60 seconds counting time. The 0.15 secs counting time is, as expected, very random but as the counting times are increased by 0.15 secs at a time, we see that this variability between the repeats is decreased. When however we reach the 0.75 secs counting time, this variability starts to rise again. (Why ??) | The most optimum window length to be used is found to be the 0.60 seconds counting time. The 0.15 secs counting time is, as expected, very random but as the counting times are increased by 0.15 secs at a time, we see that this variability between the repeats is decreased. When however we reach the 0.75 secs counting time, this variability starts to rise again. (Why ??) | ||
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Revision as of 05:17, 6 September 2007
Optimum counting time for the fluorometer
The fluorometer we are using is a Twinkle LB970 from Berthold Technologies. We noticed that by changing the counting time(window) for which the detector remains on top of each well, the variation between repeated measurements varied. We are seeking to have the less variation possible between repeats while maintaining a relatively low counting time to prevent excessive bleaching of our samples. Therefore we only examined a range of the smallest counting times possible (0.15 - 0.75 sec).
To test which is the optimum time for us to use in our experiments we followed the protocol found in the previous page. For each time point examined, different samples of the same stock solution where measured repeatedly(4 times) at different windows(counting time). From there we calculated, for each window, the percentage(%) variability between the repeated measurements(4) of the same sample. Ideally, we since the same sample is measured repeatedly, we expect the variability between the repeats to be zero(0). This however does not take into consideration the randomness inherent within fluorescence. The results are as follows:
The most optimum window length to be used is found to be the 0.60 seconds counting time. The 0.15 secs counting time is, as expected, very random but as the counting times are increased by 0.15 secs at a time, we see that this variability between the repeats is decreased. When however we reach the 0.75 secs counting time, this variability starts to rise again. (Why ??)
