J. Anim Sci.
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J. Anim Sci. 2008. 86:1291-1295. doi:10.2527/jas.2007-0498
© 2008 American Society of Animal Science
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ANIMAL GENETICS

Technical note: A novel method for routine genotyping of horse coat color gene polymorphisms1

L. J. Royo*, I. Fernández*, P. J. Azor{dagger}, I. Álvarez*, L. Pérez-Pardal* and F. Goyache*,2

* Servicio Regional de Investigación y Desarrollo Agroalimentario del Principado de Asturias-Somió, C/Camino de los Claveles 604, E-33203 Gijón (Asturias), Spain; and {dagger} Departamento de Genética, Universidad de Córdoba, Campus de Rabanales, E-14071, Córdoba, Spain

2 Corresponding author: fgoyache{at}serida.org

The aim of this note is to describe a reliable, fast, and cost-effective real-time PCR method for routine genotyping of mutations responsible for most coat color variation in horses. The melanocortin-1 receptor, Agouti-signaling peptide, and membrane-associated transporter protein alleles were simultaneously determined using 2 PCR protocols. The assay described here is an alternative method for routine genotyping of a defined number of polymorphisms. Allelic variants are detected in real time and no post-PCR manipulations are required, therefore limiting costs and possible carryover contamination. Data can be copied to a Microsoft Excel spreadsheet for semiautomatic determination of the genotype using a macro freely available at http://www.igijon.com/personales/fgoyache/software_i.htm (last accessed February 26, 2007). The performance of the method is demonstrated on 156 Spanish Purebred horses.

Key Words: horse • real-time polymerase chain reaction • coat color • melanocortin-1 receptor • Agouti-signaling peptide • membrane-associated transporter protein






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