A Simple RFLP-Based Method for HFE Gene Multiplex Amplification and Determination of Hereditary Hemochromatosis-Causing Mutation C282Y and H63D Variant with Highly Sensitive Determination of Contamination

Figure 1

Example of a compound heterozygote sample. Electrophoregram of a compound heterozygous sample (C282Y +/+, H63D +/-). (a–d) FAM-labeled PCR products (FGA STR allele), Yakima Yellow-labeled PCR products (SE33 STR alleles), and ATTO 550-labeled PCR products: FZD1 double digestion products of the FZD1 labeled on both primer size are 114 and 147 bp for SexA1 and BspHI digestion, respectively. Uncut FZD1 fragment would be 309 pb (171 or 204 bp in case of digestion failure with SexAI an BspH1, respectively). (c) ATTO 565-labeled HFE fragments digested with SexAI and BSPHI, respectively. Fragments are as labeled on the figure (H63 wild-type allele, C282Y variant, H63D variant, and C282 wild-type allele).