title: A real time PCR (RT-PCR) alternative assay to detect the T/C mutation in position 1843 of the ryanodine receptor gene.
abstract: The discovery of the causal mutation of malignant hyperthermia in pigs, a T for C substitution in base 1843 of the ryanodine receptor gene, opened the door to selection procedures based on the analysis of ryanodine receptor genotype based on PCR amplification of the region containing base 1843, subsequent digestion with specific restriction enzymes of the amplified DNA fragment, and electrophoretic analysis of the resulting bands. In this paper, we describe an assay that allows analysis of the three possible genotypes of the ryanodine receptor gene using real time PCR to amplify and detect them in a single step. Results obtained with the RT-PCR assay described in this work match 100% with those obtained using traditional PCR methods. RT-PCR methods are cheaper and faster than traditional ones allowing one to genotype up to 384 samples in a single run.
authors: Departamento de Producción Animal y Ciencia de los Alimentos, Universidad de Zaragoza, Miguel Servet 177, 50013 Zaragoza, Spain.
authors: Carmen
authors: C
authors: Burgos
authors: José Alberto
authors: JA
authors: Carrodeguas
authors: Carlos
authors: C
authors: Moreno
authors: Ana Cristina
authors: AC
authors: Sánchez
authors: Luis
authors: L
authors: Tarrafeta
authors: José Antonio
authors: JA
authors: Barcelona
authors: Pascual
authors: P
authors: López-Buesa