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Microchip Electrophoretic Analysis of Phaseolin Patterns and Its Comparison with Currently Used SDS-PAGE Techniques

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Abstract

The goal of this work was to compare reproducibility of phaseolin patterns of common bean obtained by two electrophoretic protein separation techniques including the conventional SDS-PAGE and an automated chip electrophoresis system. Five standard cultivars of common bean provided by the United States Department of Agriculture (Beltsville, Maryland) that represented five phaseolin types, T (Tendergreen), C (Contender) and S (Sanilac), B (Boyaca) and P (Pampa), were used in this study. Comparison of the phaseolin patterns revealed that the chip-on-a-lab electrophoresis provided a good reproducibility. The phaseolin polymorphism included four to seven polypeptides typical for the pattern composition of the T, C and S types. The polymorphism of the B and P patterns was also established. Phaseolin polypeptides separated by the microchip electrophoresis exhibited differences with respect to the molecular weights and electrophoretic mobility as compared to the SDS-PAGE technique. This phenomenon could be attributed to the absence of a solid separation phase in the microchip electrophoresis. Moreover, this technique has potential to substantially accelerate screening of large bean germplasm collections since it allows for the accurate analysis of the higher number of individual plants within accessions than the conventional, tedious and time consuming SDS-PAGE method.

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Acknowledgments

At the USDA-United States Department of Agriculture for ceding species Phaseolus vulgaris L. standards used in this study.

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Authors and Affiliations

  1. Banco de Germoplasma ISOPlexis, Universidade da Madeira, 9020-105, Funchal, Portugal

    Emanuel Marques da Silva, Teresa Maria Marques dos Santos, José Filipe Teixeira Ganança, Jan Jacek Slaski & Miguel Â. A. Pinheiro de Carvalho

  2. Bioresource Technologies, Alberta Innovates, Technology Futures Vegreville, Vegreville, AB, T9C 1T4, Canada

    Jan Jacek Slaski

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  1. Emanuel Marques da Silva
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  2. Teresa Maria Marques dos Santos
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  3. José Filipe Teixeira Ganança
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  5. Miguel Â. A. Pinheiro de Carvalho
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Correspondence to Emanuel Marques da Silva.

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da Silva, E.M., dos Santos, T.M.M., Ganança, J.F.T. et al. Microchip Electrophoretic Analysis of Phaseolin Patterns and Its Comparison with Currently Used SDS-PAGE Techniques. Chromatographia 76, 1163–1169 (2013). https://doi.org/10.1007/s10337-013-2511-x

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