GENETIC-MAPPING USING FLUORESCENT QUANTIFICATION OF ALLELE FREQUENCIES IN POOLED DNA LOADED BY SOLID SUPPORT

An efficient and labour-saving method for fragment analysis in linkage studies using biotinylated primers and streptavidin-coated combs is p resented. The lever of streptavidin attached to the combs was used to control the amount of immobilised material. Thus, the need for titrati on of PCR products to fit the dynamic range of the sequencer was reduc ed. The method was used to investigate the possibility of quantitating allele frequencies in pools of DNA from family members with the autos omal dominant eye disorder Best's macular dystrophy. The method allowe d the detection of one unique allele in a background of 39 other allel es. Using independent datasets, it was further found that the method w as able to detect distorted allele frequencies in affected individuals of one family as compared to reference individuals, for markers locat ed more than 30 cM from the disease locus. It was found that this proc edure is a powerful alternative to conventional linkage analysis and t he method may prove useful in a genome scan for genes involved in comp lex disorders.