The Split Hand/Split Foot (SHSF) malformation is a developmental defect of the middle rays of the hands and feet. It can occur as an isolated defect or as part of a syndrome. Reduced penetrance is frequently observed in pedigrees with SHSF. Non-syndromic and syndromic SHSF are inherited as autosomal dominant or X-linked traits and SHSF loci have been identified on chromosomes 7q and Xq. Recently the mouse dactylaplasia mutation, which appears to be analogous to SHSF, was mapped on mouse chromosome 19, an area syntenic to human chromosome 10q24-q25. We genotyped a panel of eight SHSF families for 17 markers localized to 10q24-q25. In four families multiple recombinants with several markers excluded 10q24-q25. The remaining four families provided evidence of linkage to 10q24-q25 (Zmax=3.27 at θ=0.06 for D10S677, Zmax=3.91 at θ=0 for PAX2 and Zmax=3.96 at θ=0 for D10S1239). Haplotype analysis revealed a cross-over between D10S185 and D10S677 which defined the centromeric boundary of this new SHSF locus and the telomeric boundary was defined by a cross-over between D10S1239 and D10S1237. This data places a new locus for SHSF in a 12cM region of chromosome 10q24-q25 that contains several genes including FGF8, PAX2, ZNF32 and HMX2, and confirms the genetic heterogeneity of this disorder by suggesting the presence of at least one more autosomal locus.