Team:EPF-Lausanne/Tools/MITOMI

From 2011.igem.org

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(MITOMI scans explanation)
 
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[[File:EPFL2011_MITOMIchip_beforewash_illustration.png|300px| After incubation: GFP fluorescence ]]
[[File:EPFL2011_MITOMIchip_beforewash_illustration.png|300px| After incubation: GFP fluorescence ]]
[[File:EPFL2011_MITOMIchip_beforewashDNA_illustration.png|300px| After incubation: CY5 fluorescence ]]
[[File:EPFL2011_MITOMIchip_beforewashDNA_illustration.png|300px| After incubation: CY5 fluorescence ]]
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Green fluorescence comes from the GFP-tag fused to the protein or from the incorporated Green Lysine.  
Green fluorescence comes from the GFP-tag fused to the protein or from the incorporated Green Lysine.  
Red fluorescence is due to the CY5 labeling of the DNA.
Red fluorescence is due to the CY5 labeling of the DNA.
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For data collection the fluorescence intensity measured under the button is normalised to the background and the DNA/protein ratio is calculated. This ratio is ploted against the intensity of the fluorescence of DNA in solution to make the saturation curves. And the Kd for each sequence is calculated from these curves by fitting the next equation:
For data collection the fluorescence intensity measured under the button is normalised to the background and the DNA/protein ratio is calculated. This ratio is ploted against the intensity of the fluorescence of DNA in solution to make the saturation curves. And the Kd for each sequence is calculated from these curves by fitting the next equation:
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[[File:EPFL2011_MITOMI_Kd_fiting_formula.png|150px]]
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[[File:EPFL2011_MITOMI_Kd_fiting_formula.png|180px]]
[[File:EPFL2011_MITOMI_Example_Saturation_curve_fited.png|400px]]
[[File:EPFL2011_MITOMI_Example_Saturation_curve_fited.png|400px]]

Latest revision as of 19:51, 28 October 2011