Team:Carnegie Mellon/mRFP1

From 2013.igem.org

(Difference between revisions)

Revision as of 02:30, 26 September 2013

Killer Red

mRFP1 Characterization

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.

Characterization of the Photostability of mRFP1

Photobleaching curve of mRFP1 with a HBO100 mercury-arc lamp"

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.

**Table 3** Tecan Safire II Parameters
Excitation (nm)	585
Emission (nm)	610
Excitation bandwidth (nm)	10
Emission bandwidth (nm)	10
Gain	63
Number of reads	10
Integration Time (microseconds)	40

**Table 4** Shows the fluorescence data over time during photobleaching.
Time (minutes)	Fluorescence (RFU)
0	48694
20	43083
40	36842
60	30239
80	31281
100	25273
120	21467
140	18081
160	15251
180	14427

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 <p align="center"><b> Characterization of the Photostability of mRFP1</b></p></html>
 [[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. <html><br />
+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="70%">
 <caption><p align="justify"><b>Table 3</b> Tecan Safire II Parameters</p></caption>

Team:Carnegie Mellon/mRFP1

From 2013.igem.org

Revision as of 02:30, 26 September 2013

mRFP1 Characterization

References