Materials
Castor beans Gradient former Sucrose solutions in 0.01 M EDTA, pH 7.5 33%, 44%, 50%, 57%, 60% (w/v) Grinding Medium 0.4 M sucrose 0.165 M Tris-HCl buffer, pH 7.5 0.01 M KCl 0.01 M MgCl 0.01 M EDTA adjusted to pH 7.5 0.01 M dithiothreitol Chopping board and knife or razor blade Mortar and pestle Cheesecloth Refrigerated centrifuge (ultracentrifuge preferred) with swinging bucket rotor Spectrophotometer and tubes Hemacytometer and phase contrast microscope
Procedure
1.Germinate castor beans by soaking overnight, followed by germination in moist vermiculite, or paper towels, at 30° C.
Castor beans are poisonous and may be fatal if taken internally. Wear gloves while handling.
2.After approximately 5 days, remove the embryos and cotyledons and discard. Wash the endosperm in distilled water and chill on ice.
