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Although fluorescence measurements are a fundamental tool to characterize biochemical circuits, suitable measurement instruments are usually very expensive and therefore not available for all iGEM teams. We therefore constructed a portable low-cost fluorescence detector that can be assembled for less than 15$ and can measure time traces with the sensitivity of a commercial plate reader. The assembly of the detector only requires standard electronic components and a 3D printer. We provide equations to calibrate the data from the detector including a complete consideration of the propagation of uncertainties and wrote program code to automatize data analysis. | Although fluorescence measurements are a fundamental tool to characterize biochemical circuits, suitable measurement instruments are usually very expensive and therefore not available for all iGEM teams. We therefore constructed a portable low-cost fluorescence detector that can be assembled for less than 15$ and can measure time traces with the sensitivity of a commercial plate reader. The assembly of the detector only requires standard electronic components and a 3D printer. We provide equations to calibrate the data from the detector including a complete consideration of the propagation of uncertainties and wrote program code to automatize data analysis. | ||
</p> | </p> | ||
− | + | <p> | |
+ | As a first proof of principle we measure a time trace of Cas13a digesting RNaseAlert with our detector. RNaseAlert is a commercially available molecular beacon consisting of a RNA linker, a quencher and a fluorophore, that get separated and therefore fluorescent when cut. For comparison, we also measured a time trace of the highly active RNaseA digesting RNaseAlert and a time trace of a sample containing only RNaseAlert. The time traces measured with our detector are shown in the figure below. | ||
+ | </p> | ||
+ | <div class="captionPicture"> | ||
+ | <img src="https://static.igem.org/mediawiki/2017/e/e2/T--Munich--Hardware_kinetic.png"> | ||
+ | <p> | ||
+ | Time lapse measurement of Cas13a digesting RNaseAlert on paper using our detector. The | ||
+ | positive control contains RNaseA and RNaseAlert. The negative control contains only RNaseAlert. Data points are | ||
+ | connected with lines for the convenience of the eye. Error bars represent the measurement uncertainties of the detector. | ||
+ | </p> | ||
+ | </div> | ||
</td> | </td> | ||
</tr> | </tr> |
Revision as of 10:11, 27 October 2017