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<p><h3>Characterization of chromoproteins</h3> </p> | <p><h3>Characterization of chromoproteins</h3> </p> | ||
<p style="font-family: roboto;font-size:115%;"> | <p style="font-family: roboto;font-size:115%;"> | ||
− | We found that amajLime was described as “chromoprotein” in the main page. We later found that amajLime is a fluorescent protein instead. Therefore we scanned its emission and excitation spectra and documented it in the Registry | + | We found that amajLime was described as “chromoprotein” in the main page. We later found that amajLime is a fluorescent protein instead. Therefore we scanned its emission and excitation spectra and documented it in the Registry. The excitation peak and emission peak were found to be 445 and 485 nm respectively. |
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<img src="https://static.igem.org/mediawiki/parts/1/15/Ex-Em._amajlime.png" width="50%" height="auto"> | <img src="https://static.igem.org/mediawiki/parts/1/15/Ex-Em._amajlime.png" width="50%" height="auto"> | ||
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https://static.igem.org/mediawiki/2017/9/9c/CUHK_Characterization.jpg" width="100%" height="auto"> | https://static.igem.org/mediawiki/2017/9/9c/CUHK_Characterization.jpg" width="100%" height="auto"> | ||
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− | Purified mRFP and amajLime | + | Purified mRFP and amajLime were diluted to 0.1 µg/µl (200 µl in total), in triplicates, into different buffers which pH ranges from 2 to 12. Volume of mRFP:buffer = 1:1.8, amajLime:buffer = 1:4.3. The lines represent the trend of the median fluorescence for the measured pH values. mRFP and amajLime were excited by 584 nm and 470 nm respectively and emission was measured at 607 nm and 515 nm respectively. To facilitate reproducibility of the experiment, we correlated the relative fluorescent intensity to an absolute fluorophore concentration by referring it to a standard curve of the corresponding fluorophores using the interlab study protocol (link here?). We used Rhodamine to standardize mRFP fluorescence, and Fluorescein to standardize amajLime. The results show that the optimum pH of both chromoproteins ranges from pH 6 to 8. Their fluorescent signals thus will probably not be interfered by the pH of our proposed sample: saliva or NP swab. |
Revision as of 08:08, 27 October 2017