High resolution melting analysis and sequence traces. Panel A: Normalised plot of AKT1 exon 4. Melting profiles of the wild-type control (WT) and four samples (TX25, TX94, TX285, 07M470) are shown. The melting profile of TX25 is similar to that of the wild-type control. TX94, TX285 and 07M470 show different melting profiles than the wild-type control. Panel B: Difference plot of AKT1 exon 4. Melting profiles of each sample were normalised against the wild-type control (WT). TX94, TX285 and 07M470 show distinctively different melting curves. Panel C: Sequencing of the HRM positive samples. These identified a rare SNP, IVS3-12G>A, from both TX94 and 07M470. Panel D: Sequencing identifies the E17K (c.49G>A) mutation. TX285 was positive for the E17K (c.49G>A) mutation. Sequencing was carried out in both directions to confirm the presence of the mutation.