Difference between revisions of "Part:BBa K1184000:Experience"

Revision as of 19:31, 14 September 2013

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Applications of BBa_K1184000

User Reviews

UNIQc63c6ed3490b80a8-partinfo-00000000-QINU

•••

Carnegie_Mellon 2013

Figure 1: Phototoxicity of KillerRed and mRFP1 (BBa_E1010) 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 66% ± 20% culture alive after photobleaching
KillerRed: 51% ± 17% culture alive after photobleaching

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

XL10 Ultracompetent cells were transformed with BBa_K1184000 cloned with BBa_B0034 as the RBS and BBa_R0010 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
Gain	129
Number of reads	10
Integration Time (microseconds)	40

**Table 2** Shows the fluorescence data over time during photobleaching.
Time (minutes)	Fluorescence (RFU)
0	42598
20	37616
40	33749
60	29059
80	25680
100	21985
120	19442
140	17031
160	15738
180	13741

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 </p>
 [[image:KR-Photobleach.png|thumb|600px|<b>Figure 1:</b> Photobleaching curve of KillerRed with a HBO100 mercury-arc lamp]]
-<p>XL10 Ultracompetent cells were transformed with <partinfo>BBa_K1184000</partinfo> with <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 <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 1. Fluorescence Data is shown in Table 2.</p>
 <table cellpadding="2" border="1px" cellspacing="0" align="center" width="70%">
 <caption><p align="justify"><b>Table 1</b> Tecan Safire<sup>2</sup> Parameters</p></caption>