RAPID DNA HAPLOYTYPING USING A MULTIPLEX HETERODUPLEX APPROACH - APPLICATION TO DUCHENNE MUSCULAR-DYSTROPHY CARRIER TESTING

A new strategy has been developed for rapid haplotype analysis based o n an initial multiplex amplification of several polymorphic sites, fol lowed by heteroduplex detection. Heteroduplexes formed between two dif ferent alleles are detected because they migrate differently than the corresponding homoduplexes in Hydrolink-MDE gel. This simple, rapid me thod does not depend on specific sequences such as restriction enzyme sites or CA boxes and does not require the use of isotope. This approa ch has been tested using commonly occurring polymorphisms spanning the dystrophin gene as a model, We describe the use of the method to assi gn the carrier status of females in Duchenne muscular dystrophy (DMD) pedigrees. The method may be used for other genetic diseases when muta tions are unknown or there are few dinucleotide markers in the gene pr oximity, and for the identification of haplotype backgrounds of mutant alleles. (C) 1995 Wiley-Liss, Inc.