Team:Carnegie Mellon/Project/Results
From 2013.igem.org
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<h2>KillerRed phototoxicity</h2> | <h2>KillerRed phototoxicity</h2> | ||
- | Our photobleaching experiments showed the phototoxicity of KillerRed:<br> | + | Our photobleaching experiments showed the phototoxicity of KillerRed in pSB1A3:<br> |
[[image:KillerRed RFP Phototoxicity1.png|thumb|400px|center|<b>Figure 1:</b> Phototoxicity of KillerRed and mRFP1 (<partinfo>BBa_E1010</partinfo>) on <i>E. coli</i> XL10 by irradiating with a HBO100 lamp (includes 375 nm LP filter) for 5 hours.]] | [[image:KillerRed RFP Phototoxicity1.png|thumb|400px|center|<b>Figure 1:</b> Phototoxicity of KillerRed and mRFP1 (<partinfo>BBa_E1010</partinfo>) on <i>E. coli</i> XL10 by irradiating with a HBO100 lamp (includes 375 nm LP filter) for 5 hours.]] |
Revision as of 01:04, 28 September 2013
KillerRed phototoxicity
Our photobleaching experiments showed the phototoxicity of KillerRed in pSB1A3:
KillerRed's phototoxic effect on E. coli XL10 is shown in Figure 1.
RFP 114% ± 20% (viable cells)/(Normalized RFU)
KillerRed: 53% ± 17% (viable cells)/(Normalized RFU)
KillerRed photobleaching curve
XL10 Ultracompetent cells were transformed with KillerRed (BBa_K1184000) 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 1. Fluorescence Data is shown in Table 2.
Excitation (nm) | 585 |
Emission (nm) | 610 |
Excitation bandwidth (nm) | 10 |
Emission bandwidth (nm) | 10 |
Gain | 129 |
Number of reads | 10 |
Integration Time (microseconds) | 40 |
Time (minutes) | Fluorescence (RFU) |
0 | 42598 |
20 | 37616 |
40 | 33749 |
60 | 29059 |
80 | 25680 |
100 | 21985 |
120 | 19442 |
140 | 17031 |
160 | 15738 |
180 | 13741 |