title: Adaptation of a low-cost medium-throughput genotyping system for ovine prion protein gene polymorphims associated with scrapie.
abstract: Resistance  and susceptibility to scrapie in sheep have been associated with SNPs located  at codons 136, 154 and 171 of the prion protein (PRNP) gene. Many countries  have sheep breeding programs selecting for resistance to scrapie based on the  genotyping of these SNPs. We adapted a fast and robust method for genotyping  sheep flocks for these polymorphisms, with reduced costs. Ninety-six samples  were genotyped using an adapted SNaPshot PRNP assay, and the results were  checked by resequencing. The results showed 100% concordance, using a method that  reduces genotyping costs by 70%, by reducing reagent concentrations in the  three main steps of the assay (amplicon purification, base extension and final  cleanup). This cost reduction should contribute to the development of selection  criteria based on PRNP genotyping in countries where assay costs are an  important limiting factor.
authors.affiliation: Embrapa Recursos Genéticos e Biotecnologia, Brasília, DF, Brasil. ianellabio@yahoo.com.br
authors.forename: P
authors.initials: P
authors.lastname: Ianella
authors.forename: C M
authors.initials: CM
authors.lastname: McManus
authors.forename: S R
authors.initials: SR
authors.lastname: Paiva
authors.forename: A R
authors.initials: AR
authors.lastname: Caetano
mesh_terms: D000483:Alleles; D000818:Animals; D001938:Brazil; D003062:Codon; D003365:Costs and Cost Analysis; D005838:Genotype; D060005:Genotyping Techniques / Q000191:economics / Q000662:veterinary*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D020641:Polymorphism, Single Nucleotide; D011328:Prions / Q000235:genetics*; D012608:Scrapie / Q000235:genetics*; D012756:Sheep; D034561:Sheep, Domestic
chemical_list: D003062:Codon; D011328:Prions