Most population studies of twins estimate the number of monozygotic (MZ) and dizygotic (DZ) pairs by Weinberg's differential rule. This rule assumes that within the DZ twins the numbers of unlike-sexed (U) and like-sexed (L) twins are equal. The literature on the validity of Weinberg's rule is still controversial. In this prospective population-based study (EFPTS) of 2,589 twin pairs, of whom 2,577 were of known zygosity and placentation, the estimates of Weinberg's rule agree well with the results of direct zygosity determination.