Six human minisatellite sequences (MS1, MS8, MS29, MS31, MS43, g3) have been subcloned into a stable host/vector system. Allele frequencies at the hypervariable loci detected by five of these probes were determined in a Caucasian population (200 individuals). Mendelian inheritance has been demonstrated in 5 large multi-generation pedigrees. The mutation rate has been determined in 59 families. The highest mutation rate was observed with MS1, as might be predicted from the observed high heterozygosity and in agreement with previous direct measurement of germ line mutation rates. The data presented on allele frequencies and mutation rates provide preliminary data supporting the use of these probes in paternity analysis and forensic investigations.