We have investigated the organization and genomic distribution of sequences homologous to p82H, a cloned human alpha satellite sequence purported, based on previous in situ hybridization experiments, to exist at the centromere of each human chromosome. We report here that, using Southern blotting analysis under conditions of high stringency, p82H hybridizes solely to a low-copy or single-copy alphoid domain located at or near the centromeric region of human chromosome 14. In contrast, conditions of reduced hybridization stringency permit extensive cross-hybridization with non-identical, chromosome-specific alpha satellite subsets found elsewhere in the human genome. Thus, the previously described ubiquity of "82H" human centromeric sequences reflects the existence of diverse alpha satellite subsets located at the centromeric region of each human chromosome.