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Team:Carnegie Mellon/mRFP1
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[[Image:CRQ-mRFP1.png|thumb|600px|center|The QYG-derived chromophore of the mRFP1 protein. (Z)-2-(2-(4-amino-1-imino-4-oxobutyl)-4-(4-hydroxybenzylidene)-5-oxo-4,5-dihydro-1H-imidazol-1-yl)acetic acid. The extended π-system is highlighted in red.]] | [[Image:CRQ-mRFP1.png|thumb|600px|center|The QYG-derived chromophore of the mRFP1 protein. (Z)-2-(2-(4-amino-1-imino-4-oxobutyl)-4-(4-hydroxybenzylidene)-5-oxo-4,5-dihydro-1H-imidazol-1-yl)acetic acid. The extended π-system is highlighted in red.]] | ||
<html> | <html> | ||
| + | <p align="center"><b> Characterization of the Photostability of mRFP1</b></p> | ||
| + | [[file: MRFP1.png |center|thumb|400px|''Photobleaching curve of mRFP1 with a HBO100 mercury-arc lamp"]] | ||
| + | XL10 Ultracompetent cells were transformed with <partinfo>BBa_K1184000</partinfo> cloned with <partinfo>BBa_B0034</partinfo> as the RBS and <partinfo>BBa_R0010</partinfo> as the wild-type lac promoter and induced overnight with IPTG.The overnight was bleached for 180 minutes with HBO100 (100W Mercury-arc lamp). Fluorescence data was taken using a Tecan Safire II with the parameters shown in Table 3. Fluorescence values are shown in Table 4. <br /> | ||
| + | <table cellpadding="2" border="1px" cellspacing="0" align="center" width="70%"> | ||
| + | <caption><p align="justify"><b>Table 3</b> Tecan Safire II Parameters</p></caption> | ||
| + | <tr><td><b>Excitation (nm)</b></td><td>585</td></tr> | ||
| + | <tr><td><b>Emission (nm)</b></td><td>610</td></tr> | ||
| + | <tr><td><b>Excitation bandwidth (nm)</b></td><td>10</td></tr> | ||
| + | <tr><td><b>Emission bandwidth (nm)</b></td><td>10</td></tr> | ||
| + | <tr><td><b>Gain</b></td><td>63</td></tr> | ||
| + | <tr><td><b>Number of reads</b></td><td>10</td></tr> | ||
| + | <tr><td><b>Integration Time (microseconds)</b></td><td>40</td></tr> | ||
| + | </table> | ||
| + | <br /> | ||
| + | <table cellpadding="2" border="1px" cellspacing="0" align="center" width="50%"> | ||
| + | <caption><p align="justify"><b>Table 4</b> Shows the fluorescence data over time during photobleaching.</p></caption> | ||
| + | <tr><td align="center"><b>Time (minutes)</b></td><td align="center"><b>Fluorescence (RFU)</b></td> | ||
| + | <tr><td align="left">0</td><td align="right">48694</td></tr> | ||
| + | <tr><td align="left">20</td><td align="right">43083</td></tr> | ||
| + | <tr><td align="left">40</td><td align="right">36842</td></tr> | ||
| + | <tr><td align="left">60</td><td align="right">30239</td></tr> | ||
| + | <tr><td align="left">80</td><td align="right">31281</td></tr> | ||
| + | <tr><td align="left">100</td><td align="right">25273</td></tr> | ||
| + | <tr><td align="left">120</td><td align="right">21467</td></tr> | ||
| + | <tr><td align="left">140</td><td align="right">18081</td></tr> | ||
| + | <tr><td align="left">160</td><td align="right">15251</td></tr> | ||
| + | <tr><td align="left">180</td><td align="right">14427</td></tr> | ||
| + | </table> | ||
<br> | <br> | ||
<h2>References</h2> | <h2>References</h2> | ||
</html> | </html> | ||
Revision as of 02:23, 26 September 2013
mRFP1 Characterization
Characterization of the Photostability of mRFP1
[[file: MRFP1.png |center|thumb|400px|''Photobleaching curve of mRFP1 with a HBO100 mercury-arc lamp"]] XL10 Ultracompetent cells were transformed with| Excitation (nm) | 585 |
| Emission (nm) | 610 |
| Excitation bandwidth (nm) | 10 |
| Emission bandwidth (nm) | 10 |
| Gain | 63 |
| Number of reads | 10 |
| Integration Time (microseconds) | 40 |
| Time (minutes) | Fluorescence (RFU) |
| 0 | 48694 |
| 20 | 43083 |
| 40 | 36842 |
| 60 | 30239 |
| 80 | 31281 |
| 100 | 25273 |
| 120 | 21467 |
| 140 | 18081 |
| 160 | 15251 |
| 180 | 14427 |
