Figure 3. Mutation analysis of EPHA2 in family Mu. A: Sequence trace of the wild type allele showing translation of glycine (G) at codon 948 (GGG). B: Sequence trace of the mutant allele showing the heterozygous c.2842G>T transversion (denoted K by the International Union of Pure and Applied Chemistry [IUPAC] code) that is predicted to result in the missense substitution of tryptophan (TGG) for glycine at codon 948 (p.G948W). C: Allele-specific PCR analysis using the 3 primers (Table 1) indicated by arrows in the schematic diagram; exon-17 was amplified as above with the sense (anchor) primer located in intron 16 (Ex17R1), the anti-sense primer located in the 3′-untranslated region (Ex17SF), and the nested mutant primer specific for the T-allele in codon 948 (T-alleleF). Note that only affected members of family Mu are heterozygous for the mutant T-allele (80 bp).