We have worked out a simplification of the procedure described by Schmitz et al.(Biochim. Biophys. Acta (1973) 323, 98–112) for the preparation of brush border membranes from small intestine. The procedure ultimately adopted is simple, rapid, does not necessarily require scraping and can be started from fresh or frozen material. It can be scaled up easily, allowing a quick production of large amounts of brush border membrane vesicles. These vesicles prove to be excellently suited for transport studies, as suggested by our measurements of d-glucose transport. Using these vesicles, the mode of choline transport across the brush border membrane was also investigated. Choline transport was found to occur by a saturable component with a K m of 83±4 μ M (at 20° C) and by a non-saturable component. It is independent of the presence of Na+ and appears to be nonelectrogenic.