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A Model to Determine Molecular Weights of Proteins from Gel Electrophoresis By Jose Ceja Kamyar Ghods CSUN/JPL-PAIR 2001
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Outline Getting the data (Standards) Choosing a Model Getting the data (Unknowns) Applying the model Results and conclusions
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Getting the Data Two Methods were used: Adobe Photoshop Spot Viewer
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Choosing a Model Cubic of form: log(MW)=a+b(RM)^2+c( RM)^3 Cubic of form: log(MW)=a+b(log(RM))^ 2+c(log(RM))^3 Quad Cross Validation of form: log(MW)=a+b(RM)+c(R M)^2 SLIC
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R-squared values
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Cross Validation
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Applying Our Model Collected unknown data using Photoshop Spot viewer not designed for 1D gels and not well understood. Applied best cubic model to each gel.
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Applying Our Model Created an average of our two data sets Applied cubic model to all Each standard had 3 cubic fits Used data that had the best cubic fit for each standard
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Jose’s Unknown Frog skin Gels @ 7 and 12% for males and females Within the same gel different lanes had different bands. Male and Female frog’s skin do not have the exact same proteins
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Komy’s Unknown Comparing 3 methods Overall the Manual method found the most proteins and the Amylase method found the least. The replicates of each gel were pickkin up more and different proteins.
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Conclusions & Future Work We both found that the higher concentrations found more proteins. Photoshop is more reliable for dense 1D gels. Out of the four models we tried, the cubic model was the best one. Further study is needed to find a true function relating RM to MW.
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Aknowledgements We thank CSUN/JPL-PAIR program, especially Dr. Carrol, Dr. Clevenson, Dr. Shubin, V. Hutchins and J. Handy. And our fellow students
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Residuals
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