Biomod/2011/TUM/TNT/Results: Difference between revisions

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The idea to perform bulk measurements based on FRET using a photospectrometer and a real time PCR was unsuccessful.  
The idea to perform bulk measurements based on FRET using a photospectrometer and a real time PCR was unsuccessful.  
The photospectrometer is not sensitive enough to handle Atto dyes at concentrations below 10 nM (peaks were not visible at all).  
The photospectrometer is not sensitive enough to handle Atto dyes at concentrations below 10 nM (peaks were not visible at all).  
The real time PCR, which is more sensitive, still did not deliver trustworthy data when using 50 µl samples with 10 nM Atto dyes. It could be shown that the reproducibility of the real time PCR setup was poor with deviations of up to 40 % between identical samples (figure 1) . To assure the identity of the samples a 100 µl stock was divided into two 50 µl samples. Based on these results no experiments with theU structure were performed at all with this device as the concentration of theU structure is lower than the concentration of the here test structure.  
The real time PCR, which is more sensitive, still did not deliver trustworthy data when using 50 µl samples with 10 nM Atto dyes. It could be shown that the reproducibility of the real time PCR setup was poor with deviations of up to 40 % between identical samples (figure 7). To assure the identity of the samples a 100 µl stock was divided into two 50 µl samples. Based on these results no experiments with theU structure were performed at all with this device as the concentration of theU structure is lower than the concentration of the here test structure.  


<center>[[Image:FRET efficiency Spermine skal.PNG|x250px]][[Image:FRET_efficiency_EtBr.PNG|x250px]]</center>
<center>[[Image:FRET efficiency Spermine skal.PNG|x350px|left|thumb|Fig. 7a Bulk FRET with spermine]][[Image:FRET_efficiency_EtBr.PNG|x350px|right|thumb|Fig. 7b Bulk FRET with ethidium bromide]]</center>
<font size=1>'''Figure 1: FRET efficiency Spermine and FRET efficiency EtBr'''</font>




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