Two novel real-time PCR methods for genotyping the von Willebrand disease type I mutation in Doberman Pinscher dogs.
Vet J
; 197(2): 457-60, 2013 Aug.
Article
en En
| MEDLINE
| ID: mdl-23911791
Two single tube real-time PCR methods were designed to genotype the mutation responsible for von Willebrand disease type I (von Willebrand factor c.7437G>A) in Doberman Pinscher dogs: (1) the Divergent PCR assay, which is a modification of the bi-directional PCR amplification of a specific allele (BI-PASA) technique, and (2) a minor groove binder (MGB) real-time PCR assay using fluorescently labelled probes. There was complete agreement between the genotypes determined using the two real-time PCR methods and the results of sequencing of PCR products generated by conventional PCR from genomic DNA purified from the blood of 27 Doberman Pinscher dogs. The Divergent PCR assay yielded reliable results with ≥ 6.4 ng genomic DNA per reaction and the MGB real-time PCR assay yielded reliable results with ≥ 150 pg genomic DNA per reaction. Both real-time PCR methods are suitable for routine genetic testing for the von Willebrand disease type I mutation using blood samples.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Enfermedades de von Willebrand
/
Predisposición Genética a la Enfermedad
/
Enfermedades de los Perros
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Reacción en Cadena en Tiempo Real de la Polimerasa
/
Genotipo
Límite:
Animals
Idioma:
En
Revista:
Vet J
Asunto de la revista:
MEDICINA VETERINARIA
Año:
2013
Tipo del documento:
Article
País de afiliación:
Italia
Pais de publicación:
Reino Unido