American Standardbred horses are divided into two groups based upon gait: the trot and the pace. The tendency to trot (diagonally opposite legs moving forward together) or pace (the two legs on the same side of the body moving forward together) appears to be genetically determined, although no formal genetic analysis has been undertaken. There is nearly complete assortative mating for gait; however, about 20% of the offspring sired by trotters are registered as pacers, while fewer than 1% of those sired by pacers are registered as trotters. Electrophoretically detectable genic variation at 13 protein loci has been analysed for 371 trotters and 856 pacers, and 10 blood group loci have been examined for 600 trotters and 1227 pacers. Trotters and pacers shared common alleles at 20 of the 23 loci; however, there were significant differences in allele frequencies at 21 of the 23 loci. Highly significant fixation indices (FSTS) were observed for 17 of the loci. The extent of genetic difference between Standardbred trotters and pacers was as great as or greater than that seen between some distinct horse breeds.