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Direct genotyping of the Double-Muscling locus (mh) in Belgian Blue Cattle by PCR Sequence Specific Priming (PCR-SSP)

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Author(s): Rhys H.S. Bellinge | Stephen P. A. Iaschi | Guan K. Tay

Journal: Journal of Animal and Veterinary Advances
ISSN 1680-5593

Volume: 3;
Issue: 10;
Start page: 722;
Date: 2004;
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ABSTRACT
The phenotype known as double muscling and occurring in cattle, renders breeds affected such as the Belgian Blue and Peidmontese breeds virtually cost-ineffective due to dystocia (difficult calving). Here we describe a simple PCR-based method for determining allelic heterogeneity for the common mutant allele that is based on an 11 base-pair deletion, giving rise to double muscling in the Belgian Blue breed. A set of two PCR primer pairs overlapping the deletion were designed. One pair of the set incorporated the 11 bases of the deletion site into the downstream primer, thus allowing the amplification of the wild (+11bp) allele. The primer pairs were tested on a panel of Belgian Blue and crossbred cattle that were shown to be homozygous for the wild and mutant allele as well as heterozygous crossbred containing both alleles. The use of this type of method has the potential to revolutionise the breeding industry for double muscled animals, by drawing the breeders of these animals back into the cost-effective markets. The reliance on three primers (two downstream and one common upstream primer) and polyacrylamide gel electrophoresis allows for a simple, cost effective, yet highly discriminating, DNA assay.
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