Optimization of an efficient SDS-PAGE protocol for rapid protein analysis of Brassica rapa

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Research Paper 01/08/2016
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Optimization of an efficient SDS-PAGE protocol for rapid protein analysis of Brassica rapa

Sohail Ahmad Jan, Zabta Khan Shinwari, Malik Ashiq Rabbani, Sabir Hussain Shah, Muhamad Ishaq Ibrahim, Muhammad Ilyas
J. Bio. Env. Sci.9( 2), 17-24, August 2016.
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SDS-PAGE plays a key role in the study of protein based variation among different brassica species. The present study aimed to develop an efficient Sodium Dodecyl Sulphate Poly Acrylamide Gel Electrophoresis (SDS-PAGE) protocol for B. rapa. Ten diverse genotypes were used to study their electrophoretic protein profiling. A thoroughly precise protocol was developed for optimizing the conditions such as proper pH level, centrifugation time, sample size, ammonium per sulphate (APS) concentrations, staining and de-staining time period etc. By optimizing these factors maximum polymorphic proteins were recorded that sizes range from about 10-180 k Da. All the genotypes were classified into four major groups on the basis of similarity that exist. The similarity coefficient value ranges from 40 to 95.2%. The least (40%) and maximum (95.2%) similarity coefficient values were noted among Br-508/Br-728 and Br-695/Br-725 respectively. A robust and quick SDS-PAGE protocol was developed; it will be used to study genetic diversity of other crop species and to widen the agriculture breeding program.


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