Difference between revisions of "Team:Exeter/Results"
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<h4>Experimental validation of GreenFET1J (<a href="https://2015.igem.org/Team:Exeter/Parts#toeholds">BBa_K1586000</a>):</h4> | <h4>Experimental validation of GreenFET1J (<a href="https://2015.igem.org/Team:Exeter/Parts#toeholds">BBa_K1586000</a>):</h4> | ||
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| + | <div class="slide"><a href="https://static.igem.org/mediawiki/2015/d/dd/Toehold_data.png"><img src="https://static.igem.org/mediawiki/2015/d/dd/Toehold_data.png" title="Figure 1: Table of results for BBa_K1586000 response to increasing amounts of TrigGreen."></a></div> | ||
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As was explained on the <a href="">experiments page</a>, GreenJ's (BBa_K1586000) function was characterised and validated by measuring fluorescence intensity of GFP in response to increasing amounts of trigger RNA. Shown in figure 1 is the raw data which was collected from this experiment. In order to analyse this data, the log10 of the amounts of TrigGreen (in nanograms) were calculated. So that the log10 value for 0ng could be calculated, 1*10<sup>-6</sup>ng was used instead, and all values were increased by six so that 0ng shows as 0 on the graph. These data were then plotted on a graph and a linear trend-line fitted (figure 2). On this graph, the point at about 7.72 (52.31 ng) does not match the general trend of the other points, in fact it registers at about the same fluorescence intensity as 0ng of trigger RNA. Due to the low volumes of RNA which were being added to the reactions, this reading could easily be due to a pipetting error and has therefore been left out of any further analysis. A graph without this value and with a newly fitted trend-line is shown in figure 3. As can been seen on the graph, the R<sup>2</sup> value for this is 0.7247.</br> | As was explained on the <a href="">experiments page</a>, GreenJ's (BBa_K1586000) function was characterised and validated by measuring fluorescence intensity of GFP in response to increasing amounts of trigger RNA. Shown in figure 1 is the raw data which was collected from this experiment. In order to analyse this data, the log10 of the amounts of TrigGreen (in nanograms) were calculated. So that the log10 value for 0ng could be calculated, 1*10<sup>-6</sup>ng was used instead, and all values were increased by six so that 0ng shows as 0 on the graph. These data were then plotted on a graph and a linear trend-line fitted (figure 2). On this graph, the point at about 7.72 (52.31 ng) does not match the general trend of the other points, in fact it registers at about the same fluorescence intensity as 0ng of trigger RNA. Due to the low volumes of RNA which were being added to the reactions, this reading could easily be due to a pipetting error and has therefore been left out of any further analysis. A graph without this value and with a newly fitted trend-line is shown in figure 3. As can been seen on the graph, the R<sup>2</sup> value for this is 0.7247.</br> | ||
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| + | <div class="slide"><a href="https://static.igem.org/mediawiki/2015/a/a4/Exeter_all_results_toehold_data.png"><img src="https://static.igem.org/mediawiki/2015/a/a4/Exeter_all_results_toehold_data.png" title="Figure 2: Graph of log<sub>10</sub>(TrigGreen)+6 vs. fluorescence intensity. Red point shows reading to be discarded."></a></div> | ||
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From this graph, it can be seen that the as the amount of TrigGreen increases, so does GFP fluorescence intensity, hence validating that <b>BBa_K1586000 works as expected</b>. | From this graph, it can be seen that the as the amount of TrigGreen increases, so does GFP fluorescence intensity, hence validating that <b>BBa_K1586000 works as expected</b>. | ||
Revision as of 23:05, 18 September 2015
Results
Toehold experiments
In silico testing:
Experimental validation of GreenFET1J (BBa_K1586000):
As was explained on the experiments page, GreenJ's (BBa_K1586000) function was characterised and validated by measuring fluorescence intensity of GFP in response to increasing amounts of trigger RNA. Shown in figure 1 is the raw data which was collected from this experiment. In order to analyse this data, the log10 of the amounts of TrigGreen (in nanograms) were calculated. So that the log10 value for 0ng could be calculated, 1*10-6ng was used instead, and all values were increased by six so that 0ng shows as 0 on the graph. These data were then plotted on a graph and a linear trend-line fitted (figure 2). On this graph, the point at about 7.72 (52.31 ng) does not match the general trend of the other points, in fact it registers at about the same fluorescence intensity as 0ng of trigger RNA. Due to the low volumes of RNA which were being added to the reactions, this reading could easily be due to a pipetting error and has therefore been left out of any further analysis. A graph without this value and with a newly fitted trend-line is shown in figure 3. As can been seen on the graph, the R2 value for this is 0.7247.
