Difference between revisions of "Part:BBa K1442024:Experience"
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== Experimentation == | == Experimentation == | ||
| − | <p> We compared efficiency of EMCV and NKRF in both HeLa and Huh 7.5 cells. EMCV and HeLa have been previously investigated in HeLa cells however have not been directly compared in Huh 7.5 cells. </p> | + | <p> We compared efficiency of EMCV and NKRF in both HeLa and Huh 7.5 cells. EMCV and HeLa have been previously investigated in HeLa cells however have not been directly compared in Huh 7.5 cells. We compared each of these at the peak of fluorescence for most of the cells, 1800 seconds after loading. Due to the nature of the measurement equipment we were unable to supply CO2 to the cells during measurement and the cells gradually died during the 25 hour measurement period explaining the exponential decline in fluorescence.</p> |
| − | <p></p> | + | <p><html><body><img src="https://static.igem.org/mediawiki/parts/b/b0/Flourescence_over_time_IRES_%282%29.PNG"/></body></html></p> |
| + | <p><i>This shows an exponential decline as the cells die due to lack of CO2 supply and shows the same overall trend in all the cells with maximal fluorescence occurring at approximately 1800 seconds</i></p> | ||
| + | <p></p> | ||
| + | <p>The fluorescence of each type of transfected cell was taken at 1800 seconds and averaged across the groups while fluorescence was adjusted for media and cell alone fluorescence and consolidated in this table.</p> | ||
<p><html><body><img src="https://static.igem.org/mediawiki/parts/4/49/Table_IRES.PNG"/></body></html></p> | <p><html><body><img src="https://static.igem.org/mediawiki/parts/4/49/Table_IRES.PNG"/></body></html></p> | ||
<p><i>We measured fluorescence using a tecan Magellan plate reader at wavelength 465nm - 530nm (that corresponding to GFP). Each IRES had ten replicates in each cell and hence error bars had an accuracy of 0.1. The fluorescence of media and cells themselves were accounted for in each case and the negative control used was the human optimised forwards GFP <html><body><a href="https://parts.igem.org/Part:BBa_K1442106">part:BBa_K1442106</a></body></html> without IRES included.</i></p> | <p><i>We measured fluorescence using a tecan Magellan plate reader at wavelength 465nm - 530nm (that corresponding to GFP). Each IRES had ten replicates in each cell and hence error bars had an accuracy of 0.1. The fluorescence of media and cells themselves were accounted for in each case and the negative control used was the human optimised forwards GFP <html><body><a href="https://parts.igem.org/Part:BBa_K1442106">part:BBa_K1442106</a></body></html> without IRES included.</i></p> | ||
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<p><html><body><img src="https://static.igem.org/mediawiki/parts/0/05/Bar_chart_all_IRES_with_error_bars.PNG"/></body></html> | <p><html><body><img src="https://static.igem.org/mediawiki/parts/0/05/Bar_chart_all_IRES_with_error_bars.PNG"/></body></html> | ||
Revision as of 15:51, 11 October 2014
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Experimentation
We compared efficiency of EMCV and NKRF in both HeLa and Huh 7.5 cells. EMCV and HeLa have been previously investigated in HeLa cells however have not been directly compared in Huh 7.5 cells. We compared each of these at the peak of fluorescence for most of the cells, 1800 seconds after loading. Due to the nature of the measurement equipment we were unable to supply CO2 to the cells during measurement and the cells gradually died during the 25 hour measurement period explaining the exponential decline in fluorescence.
This shows an exponential decline as the cells die due to lack of CO2 supply and shows the same overall trend in all the cells with maximal fluorescence occurring at approximately 1800 seconds
The fluorescence of each type of transfected cell was taken at 1800 seconds and averaged across the groups while fluorescence was adjusted for media and cell alone fluorescence and consolidated in this table.
We measured fluorescence using a tecan Magellan plate reader at wavelength 465nm - 530nm (that corresponding to GFP). Each IRES had ten replicates in each cell and hence error bars had an accuracy of 0.1. The fluorescence of media and cells themselves were accounted for in each case and the negative control used was the human optimised forwards GFP part:BBa_K1442106 without IRES included.
UNIQ8bc46474c78d679e-partinfo-00000004-QINU
UNIQ8bc46474c78d679e-partinfo-00000005-QINU