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.
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).
Nakajima
E
E
Animal Genome Research Program Team, STAFF Institute, Ibaraki, Japan.
Matsumoto
T
T
Yamada
R
R
Kawakami
K
K
Takeda
K
K
Ohnishi
A
A
Komatsu
M
M
D000818:Animals; D001483:Base Sequence; D015220:Calcium Channels / Q000235:genetics*; D004247:DNA / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D005260:Female; D005796:Genes / Q000235:genetics*; D005838:Genotype; D008297:Male; D009124:Muscle Proteins / Q000235:genetics*; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012150:Polymorphism, Restriction Fragment Length; D018807:Polymorphism, Single-Stranded Conformational; D019837:Ryanodine Receptor Calcium Release Channel; D013552:Swine / Q000235:genetics*
D015220:Calcium Channels; D009124:Muscle Proteins; D019837:Ryanodine Receptor Calcium Release Channel; D004247:DNA