Random spore analysis of crosses between a strain bearing the ade6-M26 hotspot mutation and strains bearing other ade6 mutations was performed. Recombinant prototroph frequencies increase with increasing distance from M26 for mutations both 5' and 3' of M26. Maximum prototroph frequencies are obtained for mutations lying more than 700 nucleotides downstream from M26. Similar results are obtained for crosses with the ade6-M375 control mutation, but the prototroph frequencies are lower. The factor of stimulation of recombination by M26 as compared to the M375 control (M26 marker effect) also displays distance-dependence. These results are discussed in the context of the mechanism of M26 recombination, as well as in relation to recombination initiation, hybrid DNA formation, and mismatch repair at ade6. Keywords Conversion middle dot M26 hotspot middle dot Recombination middle dot Schizosaccharomyces pombe