J. Anim Sci.
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Journal of Animal Science, Vol 74, Issue 12 2904-2906, Copyright © 1996 by American Society of Animal Science

JOURNAL ARTICLE

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

E. Nakajima, T. Matsumoto, R. Yamada, K. Kawakami, K. Takeda, A. Ohnishi and M. Komatsu
Animal Genome Research Program Team, STAFF Institute, Ibaraki, Japan.

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).




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Copyright © 1996 by the American Society of Animal Science.