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Rapid DNA haplotyping using a multiplex heteroduplex approach: Application to duchenne muscular dystrophy carrier testing
Author(s) -
Prior Thomas W.,
Wenger Gail D.,
Papp Audrey C.,
Snyder Pamela J.,
Sedra Mary S.,
Bartolo Claire,
Moore Jay W.,
Highsmith W. Edward
Publication year - 1995
Publication title -
human mutation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 162
eISSN - 1098-1004
pISSN - 1059-7794
DOI - 10.1002/humu.1380050312
Subject(s) - heteroduplex , haplotype , biology , genetics , duchenne muscular dystrophy , dystrophin , multiplex , multiplex polymerase chain reaction , allele , restriction enzyme , muscular dystrophy , pedigree chart , gene , computational biology , polymerase chain reaction
A new strategy has been developed for rapid haplotype analysis based on an initial multiplex amplification of several polymorphic sites, followed by heteroduplex detection. Heteroduplexes formed between two different alleles are detected because they migrate differently than the corresponding homoduplexes in Hydrolink‐MDE gel. This simple, rapid method does not depend on specific sequences such as restriction enzyme sites or CA boxes and does not require the use of isotope. This approach has been tested using commonly occurring polymorphisms spanning the dystrophin gene as a model. We describe the use of the method to assign the carrier status of females in Duchenne muscular dystrophy (DMD) pedigrees. The method may be used for other genetic diseases when mutations are unknown or there are few dinucleotide markers in the gene proximity, and for the identification of haplotype backgrounds of mutant alleles. © 1995 Wiley‐Liss, Inc.