Team:Carnegie Mellon/Project/Results

From 2013.igem.org

(Difference between revisions)
Line 3: Line 3:
<h1>Experimental results</h1>
<h1>Experimental results</h1>
-
[[image:KillerRed RFP Phototoxicity1.png|thumb|600px|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.]]
<br><br>
<br><br>
<p>KillerRed's phototoxic effect on <i>E. coli</i> XL10 is shown in Figure 1.<br />
<p>KillerRed's phototoxic effect on <i>E. coli</i> XL10 is shown in Figure 1.<br />
Line 9: Line 9:
KillerRed: 53% ± 17% (viable cells)/(Normalized RFU)
KillerRed: 53% ± 17% (viable cells)/(Normalized RFU)
</p><br><br>
</p><br><br>
-
[[image:KR-Photobleach.png|thumb|600px|center|<b>Figure 1:</b> Photobleaching curve of KillerRed with a HBO100 mercury-arc lamp]]
+
[[image:KR-Photobleach.png|thumb|400px|center|<b>Figure 1:</b> Photobleaching curve of KillerRed with a HBO100 mercury-arc lamp]]
<br><br>
<br><br>
<p>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.</p>
<p>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.</p>

Revision as of 00:54, 28 September 2013

Killer Red





Experimental results

Figure 1: Phototoxicity of KillerRed and mRFP1 (<partinfo>BBa_E1010</partinfo>) on E. coli XL10 by irradiating with a HBO100 lamp (includes 375 nm LP filter) for 5 hours.



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)



Figure 1: Photobleaching curve of KillerRed with a HBO100 mercury-arc lamp



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.





Table 1 Tecan Safire2 Parameters
Excitation (nm)585
Emission (nm)610
Excitation bandwidth (nm)10
Emission bandwidth (nm)10
Gain129
Number of reads10
Integration Time (microseconds)40

Table 2 Shows the fluorescence data over time during photobleaching.
Time (minutes)Fluorescence (RFU)
042598
2037616
4033749
6029059
8025680
10021985
12019442
14017031
16015738
18013741



Retrieved from "http://2013.igem.org/Team:Carnegie_Mellon/Project/Results"