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Team:Carnegie Mellon/mRFP1
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| - | <h2>mRFP1 | + | <h2>mRFP1 characterization</h2> |
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| - | [[Image:CRQ-mRFP1.png|thumb| | + | [[Image:CRQ-mRFP1.png|thumb|300px|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.]] |
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| - | <p align="center"><b> Characterization of the Photostability of mRFP1</b></p> | + | <p align="center"><b> Characterization of the Photostability of mRFP1</b></p></html> |
| - | [[ | + | [[Image:MRFP1.png|center|thumb|400px|Photobleaching curve of mRFP1 with a HBO100 mercury-arc lamp]] |
| - | XL10 Ultracompetent cells were transformed with <partinfo> | + | XL10 Ultracompetent cells were transformed with <partinfo>BBa_E1010</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. <html><br /> |
| - | <table cellpadding="2" border="1px" cellspacing="0" align="center" width=" | + | <table cellpadding="2" border="1px" cellspacing="0" align="center" width="40%"> |
<caption><p align="justify"><b>Table 3</b> Tecan Safire II Parameters</p></caption> | <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>Excitation (nm)</b></td><td>585</td></tr> | ||
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| - | <table cellpadding="2" border="1px" cellspacing="0" align="center" width=" | + | <table cellpadding="2" border="1px" cellspacing="0" align="center" width="40%"> |
<caption><p align="justify"><b>Table 4</b> Shows the fluorescence data over time during photobleaching.</p></caption> | <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="center"><b>Time (minutes)</b></td><td align="center"><b>Fluorescence (RFU)</b></td> | ||
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<h2>References</h2> | <h2>References</h2> | ||
| + | Campbell,R.E., Tour,O., Palmer,A.E., Steinbach,P.A., Baird,G.S., Zacharias,D.A. and Tsien,R.Y. <i>A monomeric red fluorescent protein</i>. Proc. Natl. Acad. Sci. U.S.A. 99 (12), 7877-7882 (2002) | ||
</html> | </html> | ||
Latest revision as of 01:21, 28 September 2013
mRFP1 characterization
Characterization of the Photostability of mRFP1
XL10 Ultracompetent cells were transformed with <partinfo>BBa_E1010</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.
| 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 |
