In the vertebrate retina, rods mediate twilight vision and cones daylight vision. Rods have been purified easily from the retina, and thus the phototransduction mechanism in rods is now well documented. However, it has not been possible to purify cones in large quantities, and therefore, the knowledge on the mechanism in cones is limited. Here we report purification of carp (Cyprinus carpio) cones with a stepwise Percoll gradient. Using purified cells, we compared the phototransduction mechanism between rods and cones. The results showed that both transducin activation and phosphodiesterase activation are less effective, and visual pigment phosphorylation is faster in cones. These differences explain lower light-sensitivity and briefer photoresponse time course in cones.