Fig. 1

DNA sequence of codons 153–156 in the canine APC gene determined by PCR-direct sequencing and schemes of PCR-RFLP and TaqMan duplex real-time PCR assays for the canine APC genotyping. a PCR-direct sequencing. Representative results of peripheral blood-derived DNAs of JRTs with and without the germline APC variants (right and left panels). Note heterozygous deletion-insertion variant at codons 154 and 155 in a carrier dog (arrows). b PCR-RFLP assay. Note that, while the 156-bp fragment of the wild-type APC allele contains two restriction sites of MseI, there are 3 restriction sites in the fragment of the mutant APC allele. Restriction sites of MseI (T/TAA) are highlighted in blue. Asterisks indicate the variant sites. Uppercase and lowercase letters indicate exon 4 and introns, respectively. c TaqMan duplex real-time PCR assay. Probes specific for wild-type and mutant alleles are highlighted in yellow and green, respectively. Asterisks indicate variant sites