Team:Carnegie Mellon/Project/Results

From 2013.igem.org

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[[file:KR-Photobleach.png|thumb|600px|center|<b>Figure 1:</b> Photobleaching curve of KillerRed with a HBO100 mercury-arc lamp]]
[[file:KR-Photobleach.png|thumb|600px|center|<b>Figure 1:</b> Photobleaching curve of KillerRed with a HBO100 mercury-arc lamp]]
<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>
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<table cellpadding="2" border="1px" cellspacing="0" align="center" width="50%">
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<table cellpadding="2" border="1px" cellspacing="0" align="center" width="40%">
<caption><p align="justify"><b>Table 1</b> Tecan Safire<sup>2</sup> Parameters</p></caption>
<caption><p align="justify"><b>Table 1</b> Tecan Safire<sup>2</sup> 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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<tr><td><b>Integration Time (microseconds)</b></td><td>40</td></tr>
<tr><td><b>Integration Time (microseconds)</b></td><td>40</td></tr>
</table>
</table>
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<table cellpadding="2" border="1px" cellspacing="0" align="center" width="50%">
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<table cellpadding="2" border="1px" cellspacing="0" align="center" width="40%">
<caption><p align="justify"><b>Table 2</b> Shows the fluorescence data over time during photobleaching.</p></caption>
<caption><p align="justify"><b>Table 2</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>

Revision as of 22:00, 26 September 2013

Killer Red



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