Numerous case-control studies have addressed the hypothesis that variant alleles of the dopamine D2 receptor gene (DRD2) increase the liability for alcohol and/or drug dependence, and both positive and negative results have been reported. Because population frequencies of these alleles vary considerably, the conflicting results could be due to population stratification bias. Using the transmission disequilibrium test, the present study examined linkage disequilibrium of alcohol and drug (opioid and/or cocaine) dependence with three DRD2 polymorphic systems: (a) TaqI A, (b) TaqI D, and (c) the functional -141CIns/Del promoter systems. DNA samples were collected from small nuclear families (SNFs), where one or more offspring met DSM-III-R or DSM-IV criteria for alcohol and/or drug dependence. Because positive association between DRD2 alleles and alcohol and/or drug dependence has been reported only in populations of European ancestry, we limited the present study to European Americans (EAs). No evidence for linkage disequilibrium was found for any of the polymorphic systems when examined in relation to any substance dependence, alcohol dependence (with or without drug dependence), or drug dependence (with or without alcohol dependence). These results are consistent with those from a recent family-based study of alcohol dependence. Together, these studies suggest that the conflicting findings from case-control studies of the association between alleles of DRD2 and substance dependence may be attributable to population stratification in some samples.