Table 3.
Reaction Primer Mixtures and Expected PCR Products
| Reaction | Allele-specific primer and label | Reverse primer | PCR control primer set | Predicted PCR products |
| 282C-allele (WT) specific | C282Y-WT (37.5pM) CYA-ROX (Red) | 223-RVR (12.5pM) | HFECONFWD (12.5pM) 363-RVR(12.5pM) | 223 bp (red) C-product 363 bp (red) PCR control |
| 282Y-allele (VT) specific | C282Y-VT (34pM) CYA-R110 (Blue) | 232-RVR (12.5pM) | HFECONFWD (18.8pM) 528-RVR (18.8pM) | 232 bp (blue) Y-product 528 bp (red) PCR control |
| 63H-allele (WT) specific | H63D-WT (37.5pM) CYA-ROX (Red) | 174-RVR (12.5pM) | HFECONFWD (10pM) 293-RVR (10pM) | 174 bp (red) H-product 293 bp (red) PCR control |
| 63D-allele (VT) specific | H63D-VT (37.5pM) CYA-R110 (Blue) | 160-RVR (12.5pM) | HFECONFWD (9.4pM) 244-RVR (9.4pM) | 160 bp (blue) D-product 244 bp (red) PCR control |
| 65S-allele (WT) specific | S65C-WT (31.3pM) CYA-ROX (Red) | 201-RVR (12.5pM) | HFECONFWD (6.3pM) 268-RVR (6.3pM) | 211 bp (red) S-product 268 bp (red) PCR control |
| 65C-allele (VT) specific | S65C-VT (31.3pM) CYA-R110 (Blue) | 211-RVR (12.5pM) | HFECONFWD (9.4pM) 281-RVR (9.4pM) | 201 bp (blue) C-produce 281 bp (red) PCR control |
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↵The PCR control product is red-labeled when the acceptor dye on the HFECONFWD primer is ROX, and black-labeled when it is TAM.
