Technical note: use of a PCR-single strand conformation polymorphism (PCR-SSCP) for detection of a point mutation in the swine ryanodine receptor (RYR1) gene.

Abstract

We used the polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis for screening the point mutation (C1843 to T) in the porcine ryanodine receptor (RYR1) gene. The PCR products (659 bp) were heat-denatured and separated by polyacrylamide gel electrophoresis. On silver-stained gels, the point mutation within the RYR1 gene could be detected clearly by mobility shifts. The best conditions for detecting the point mutation were by using a 5% polyacrylamide gel without glycerol and loading at 3 degrees C. The RYR1 genotypes diagnosed by PCR-SSCP were identical to the genotypes diagnosed by restriction enzyme fragment length polymorphism in all cases examined (n = 606).