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of the second PCR products was incubated for 2 h at 37 °C in a volume of 15 μL using 2 U of a restriction enzyme and subsequently resolved by 5% denaturing polyacrylamide gel electrophoresis (PAGE) with silver staining according to the procedure of

result of the high resolution ratio and repeatability, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) is being widely used in molecular marker research for many different crops. However, SDS-PAGE cannot reveal much genetic

annealing temperature because of the different primers used. The amplification products were separated using 8% polyacrylamide gel electrophoresis at 75 W (Bio-Rad Electrophoresis system, Hercules, CA) for 2 h and then visualized using a simplified silver

/volume) agarose gel containing ethidium bromide 1.0 μg·mL –1 in 1 × TAE (40 m m Tris-acetate, 1 m m EDTA) buffer, observed under ultraviolet light and photographed. SSR PCR products were separated by polyacrylamide gel electrophoresis (6% gel) and silver

antioxidant enzyme. The extracts (15 μL) for SOD, CAT, and POD were loaded on each gel. Native polyacrylamide gel electrophoresis (PAGE) was performed using a Mini-Protean system (Bio-Rad Laboratories, Hercules, CA) at 4 °C, 120 V for 90 min, except that

/v) polyvinylpyrrolidone. Protein concentration was determined as described in Bardford (1976) using bovine serum albumin as the standard. The separation of total proteins was performed using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) in 12

45 s at 72 °C, and a final extension for 7 min at 72 °C. PCR amplifications were carried out in a GeneAmp PCR system 9700 (Applied Biosystems, Foster City, CA). The resulting products were subjected to 8% polyacrylamide gel electrophoresis at 160 V

3–10], and the concentration was determined according to Bradford (1976) using bovine serum albumin as standard. Protein separation and image analysis. Proteins were separated by two-dimensional polyacrylamide gel electrophoresis (2D

volume of 1 mL, as described by Fernández-García et al. (2004) . The mean value of at least three replications was taken as indicator of peroxidase activity. Determination of isoperoxidase profile by native polyacrylamide gel electrophoresis (PAGE

Mol. Biol. Rpt. 13 207 209 Ghosh, A.K. Lukens, L.N. Hunter, D.M. Strommer, J.N. 2006 European and asian pears: Simple sequence repeat-polyacrylamide gel electrophoresis-based analysis of