Technical note: a novel method for routine genotyping of horse coat color gene polymorphisms.
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.
Royo
L J
LJ
Servicio Regional de Investigación y Desarrollo Agroalimentario del Principado de Asturias-Somió, C/Camino de los Claveles 604, E-33203 Gijón (Asturias), Spain.
Fernández
I
I
Azor
P J
PJ
Alvarez
I
I
Pérez-Pardal
L
L
Goyache
F
F
D054366:Agouti Signaling Protein / Q000235:genetics*; D000483:Alleles; D000818:Animals; D017931:DNA Primers; D015342:DNA Probes; D005260:Female; D005838:Genotype; D006200:Hair Color / Q000235:genetics*; D006736:Horses / Q000235:genetics*; D008297:Male; D026901:Membrane Transport Proteins / Q000235:genetics*; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods / Q000592:standards / Q000662:veterinary; D044102:Receptor, Melanocortin, Type 1 / Q000235:genetics*; D012680:Sensitivity and Specificity
D054366:Agouti Signaling Protein; D017931:DNA Primers; D015342:DNA Probes; D026901:Membrane Transport Proteins; D044102:Receptor, Melanocortin, Type 1