Fig. 3

a. DNA sequencing electrophoretograms of the CUBN exon 55/intron 55 boundary. Shown are results of a normal control (above) and a Komondor dog with hereditary selective intestinal cobalamin malabsorption (below). The cartoon indicates the 3′ end of exon 55 (box) and the 5′ end of intron 55 (line) aligned with the sequences. The splice donor consensus has a solid underline in the normal dog sequence. The CUBN c.NM_001003148.1; c.8746 + 1G > A variant is indicated by the black triangle and a potential cryptic splice donor has a dotted underline in the affected dog sequence. b. Genotyping of the CUBN c.NM_001003148.1; c.8746 + 1G > A splicing variant in a Komondor family segregating I-GS. Genomic DNA samples were amplified by PCR using primers flanking the G/A variant of the intron 55 splice donor consensus (Table 1, Additional file 2). Bce AI digestion fragments of the PCR amplicons were separated on 4% agarose gels. An endonuclease control cut-site created 293 and 68 bp fragments from the 361 bp amplicon. Cutting at the wildtype G recognition site further cut the 293 bp fragment into 237 and 56 bp fragments. The deduced genotypes are shown below each lane, and symbols above indicate the sex and disease status of each dog as in Fig. 2