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− | + | <h2 class="section-heading">Calibrations</h2> | |
− | + | <p class="lead"> Before our measurements began, we performed some calibrations: First an OD<sub>600</sub> reference point for our plate reader, performed with LUDOX according to the protocol. Here we found a correction factor which can be used to calculate OD from measured absorbance. Our correction fator is 3.11. <br><br> | |
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− | + | Secondly we made a fluorescence standard curve with a serial dilution of fluorescein (figure 1). We used the lower 5 data points to calculate a mean µM fluorescein pr a.u. We chose to use the lower concentration range due to two factors: 1) Linearity is better for the lower fluorescein concentrations, and 2) our measured data has a maximum fluorescence of 500, which makes it more important to have a good fit in the lower range. | |
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+ | <figcaption><b>Figure 1 </b>Standard curve of fluorescein fluorescence. | ||
+ | Fluorescence in arbitraty units (a.u.), fluorescein concentration in µM.</figcaption> | ||
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− | + | <h2 class="section-heading">Calibrations</h2> | |
− | < | + | <p class="lead"> Before our measurements began, we performed some calibrations: First an OD<sub>600</sub> reference point for our plate reader, performed with LUDOX according to the protocol. Here we found a correction factor which can be used to calculate OD from measured absorbance. Our correction fator is 3.11. <br><br> |
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− | <br> | + | Secondly we made a fluorescence standard curve with a serial dilution of fluorescein (figure 1). We used the lower 5 data points to calculate a mean µM fluorescein pr a.u. We chose to use the lower concentration range due to two factors: 1) Linearity is better for the lower fluorescein concentrations, and 2) our measured data has a maximum fluorescence of 500, which makes it more important to have a good fit in the lower range. |
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+ | <img class="img-responsive" src="https://static.igem.org/mediawiki/2017/b/b9/InterLab-figure_UCopenhagen.png" alt="" width="250" height="200"> | ||
+ | |||
+ | <figcaption><b>Figure 1 </b>Standard curve of fluorescein fluorescence. | ||
+ | Fluorescence in arbitraty units (a.u.), fluorescein concentration in µM.</figcaption> | ||
+ | </figure> | ||
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Revision as of 17:59, 29 October 2017
Calibrations
Before our measurements began, we performed some calibrations: First an OD600 reference point for our plate reader, performed with LUDOX according to the protocol. Here we found a correction factor which can be used to calculate OD from measured absorbance. Our correction fator is 3.11.
Secondly we made a fluorescence standard curve with a serial dilution of fluorescein (figure 1). We used the lower 5 data points to calculate a mean µM fluorescein pr a.u. We chose to use the lower concentration range due to two factors: 1) Linearity is better for the lower fluorescein concentrations, and 2) our measured data has a maximum fluorescence of 500, which makes it more important to have a good fit in the lower range.
Calibrations
Before our measurements began, we performed some calibrations: First an OD600 reference point for our plate reader, performed with LUDOX according to the protocol. Here we found a correction factor which can be used to calculate OD from measured absorbance. Our correction fator is 3.11.
Secondly we made a fluorescence standard curve with a serial dilution of fluorescein (figure 1). We used the lower 5 data points to calculate a mean µM fluorescein pr a.u. We chose to use the lower concentration range due to two factors: 1) Linearity is better for the lower fluorescein concentrations, and 2) our measured data has a maximum fluorescence of 500, which makes it more important to have a good fit in the lower range.