Figure 3

Assay design (A) and example of the results (B) obtained with polymerase chain reaction (PCR)-restriction fragment length polymorphism method. A 203 bp fragment TasR38 gene that covers the first SNP site of interest was amplified and subsequently digested with HaeIII restriction enzyme that recognizes only the C allele (a). A 194 bp fragment TasR38 gene that covers the second and third SNP sites of interest was amplified and subsequently digested with Eco47III restriction enzyme that recognizes only the C allele and with RsaI that recognizes the G allele. The resulting fragments that show the genotype of each individual were analyzed on polyacrylamide gel in parallel with the untreated PCR products. a M, size marker; G/G, C/G, and C/C correspond to the SNP alleles of polymorphism rs713598. The arrows indicate the DNA fragments (203, 156 bp) obtained after HaeIII restriction enzyme digestion. A 47 bp restriction fragment was eluted from the gel. b M, size marker; on the left of the size marker, C/C, T/T, and C/T correspond to the SNP alleles of polymorphism rs1726866; on the right of the size marker, G/G, G/A, and A/A correspond to the SNP alleles of polymorphism rs10246939. The arrows indicate the DNA fragments (194, 149 bp) obtained after Eco47III and RsaI restriction enzyme digestion. A 45 bp restriction fragment was eluted from the gel. The gel was stained with ethidium bromide.