Development of a snapback method of single-strand conformation polymorphism analysis for genotyping Golden Retrievers for the X-linked muscular dystrophy allele

Honeyman, K., Carville, K.S., Howell, J.M., Fletcher, S. and Wilton, S.D. (1999) Development of a snapback method of single-strand conformation polymorphism analysis for genotyping Golden Retrievers for the X-linked muscular dystrophy allele. American Journal of Veterinary Research, 60 (6). pp. 734-7.

Abstract

OBJECTIVE

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

ANIMALS

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

PROCEDURE

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.

CONCLUSION AND CLINICAL RELEVANCE

Snapback SSCP analysis provided a simple, fast, and accurate method for genotyping Golden Retrievers for the mutation known to cause X-linked muscular dystrophy.

Item Type:	Journal Article
Murdoch Affiliation(s):	School of Veterinary and Biomedical Sciences
Publisher:	American Veterinary Medical Association
URI:	http://researchrepository.murdoch.edu.au/id/eprint/17880

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