Optimization of an efficient SDS-PAGE protocol for rapid protein analysis of Brassica rapa
By: Sohail Ahmad Jan, Zabta Khan Shinwari, Malik Ashiq Rabbani, Sabir Hussain Shah, Muhamad Ishaq Ibrahim, Muhammad Ilyas
Key Words: Brassica rapa, SDS-PAGE, Genetic variation, Phylogenetic relationship.
J. Bio. Env. Sci. 9(2), 17-24, August 2016.
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.