Development of a snapback method of single-strand conformation polymorphism analysis for genotyping Golden Retrievers for the X-linked muscular dystrophy allele.
OBJECTIVE
To develop a snapback method of single-strand conformation polymorphism (SSCP) analysis for genotyping Golden Retrievers for the X-linked muscular dystrophy allele.


METHODS
20 Golden Retriever puppies from a colony with X-linked muscular dystrophy.


METHODS
DNA spanning the canine dystrophin mutation was amplified by means of a polymerase chain reaction (PCR), using a primer modified to have an additional sequence at the 5' terminus. The primer was designed so that 1 terminus of the single-stranded PCR product could anneal to the normal sequence flanking the region of the mutation in the allele but not in the mutant allele. True disease status of the dogs was determined by means of a PCR and restriction digest protocol.


RESULTS
Snapback SSCP analysis allowed for accurate and unambiguous genotyping of unaffected, carrier, and affected dogs, whereas conventional SSCP analysis, using the unmodified primer, did not. Creatine kinase activities measured within 24 hours after birth were not consistent with genotype.


CONCLUSIONS
Snapback SSCP analysis provided a simple, fast, and accurate method for genotyping Golden Retrievers for the mutation known to cause X-linked muscular dystrophy.
Honeyman
K
K
Department of Pathology, University of Western Australia, Australian Neuromuscular Research Institute, QEII Medical Centre, Nedlands.
Carville
K S
KS
Howell
J M
JM
Fletcher
S
S
Wilton
S D
SD
D000818:Animals; D000831:Animals, Newborn; D001483:Base Sequence; D003402:Creatine Kinase / Q000097:blood; D004283:Dog Diseases / Q000097:blood / Q000235:genetics*; D004285:Dogs; D005091:Exons; D005260:Female; D005838:Genotype; D007438:Introns; D008297:Male; D008969:Molecular Sequence Data; D009137:Muscular Dystrophy, Animal / Q000097:blood / Q000235:genetics*; D010375:Pedigree; D018807:Polymorphism, Single-Stranded Conformational; D014960:X Chromosome
D003402:Creatine Kinase