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

¹ SERIDA-Somió, C/ Camino de los Claveles 604, E-33203 Gijón (Asturias), Spain
² Departamento de Genética, Universidad de Córdoba, Campus de Rabanales, E-14071-Córdoba, Spain

^* To whom correspondence should be addressed. E-mail: fgoyache{at}serida.org.

	Abstract

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. MC1R, ASIP, and MATP alleles were simultaneously determined using 2 different PCR protocols. The assay described here is an alternative method for routine typing 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 carry-over contamination. Data can be copied to a Microsoft Excel-spreadsheet for semi-automatic determination of the genotype using a macro freely available at http://www.igijon.com/personales/fgoyache/software_i.htm. The performance of the method is demonstrated on 156 Spanish Purebred horses.

Key Words: horse, RT-PCR, coat color, MC1R, ASIP, MATP