Evaluation location: Iowa, United States
Materials and methods followed that of Cronn et al, TAG(1997) 95:532-545. Starch gel electrophoresis was performed on crude protein extracts of cotyledon tissue. Approximately 20 plants were analyzed per accession (number of plants is recorded in sample size). Tissue was homoginized in a 1.5 ml centrifuge tube with a power-driven acetal pestle (on ice) in solubilization buffer [75 mM sodium phosphate (pH 7.5), 100 mM sodium ascorbate, 10 mM sodium diethyldithiocarbamate, 10 mM dithioerythritol, 10% (w/v) PVP-40, 5% sucrose, 0.5% bovine serum albumin and 10 mM B-mercaptoethanol]. Extracts were frozen at -70 C until use. Enzymes were separated on 12% starch gels and were visualized. Refer to Cronn et al. for allele and locus designation.