Determination of HMW - GS in wheat using SDS - PAGE and Lab-on-chip methods
Keywords:wheat, HMW-GS, SDS-PAGE, LoC
SDS-PAGE is widely used to determine the amounts of the different gluten protein types. However, this method is time-consuming, especially at early stages of wheat breeding, when large number of samples needs to be analyzed. On the other hand, LoC (Lab-on-Chip) technique has the potential for a fast, reliable, and automatable analysis of proteins. Benefits and limitations of Lab-on-Chip method over SDS-PAGE method in gluten proteins evaluation were explored in order to determine in which way LoC method should be improved in order to make its results more compliant with the results of SDS-PAGE. Chip electrophoresis provides a very good reproducibility of HMW-GS patterns. Moreover this approach is much faster than the conventional SDS-PAGE methods requiring several hours for an analysis. Another advantage over traditional gel electrophoresis is lower sample and reagent volume requirements, as well as specialized protein standards for accurate reproducibility and quantification. In the present study, we identified novel complex allele located at the locus Glu-1B.
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