Table 2.
Primer sequences
| Primer name | Sequence 5′-3′ | Function |
| C282Y-WT | CTGGGTGCTCCACCTGGC | ASP for C282Y C allele |
| C282Y-VT | CTGGGTGCTCCACCTGGT | ASP for C282Y Y allele |
| H63D-WT | TCCACACGGCGACTCTCATG | ASP for H63D H allele |
| H63D-VT | TCCACACGGCGACTCTCATC | ASP for H63D D allele |
| S65C-WT | AGTTCGGGGCTCCACACGGCGACT | ASP for S65C S allele |
| S65C-VT | AGTTCGGGGCTCCACACGGCGACA | ASP for S65C C allele |
| HFECONFWD | CGGTGGTCCGGTGACCAGAGATGCAGCG | Forward PCR control primer |
| 160-RVR | ACATGGTTAAGGCCTGTTGC | Forward H63D VT primer |
| 174-RVR | TCCCCTCCTACTACACATGG | Forward H63D WT primer |
| 211-RVR | AGGTGTGTGGAGCCTCAACA | Forward S65C WT primer |
| 201-RVR | AGCCTCAACATCCTGCTCCC | Forward S65C VT primer |
| 223-RVR | ATCATGTGACCTCTTCAGTGACC | Forward C282Y WT primer |
| 232-RVR | AAGGTGACACATCATGTGACC | Forward C282Y VT primer |
| 244-RVR | CTTGACTCAATGATTATCAGAGTGG | Reverse PCR control primer |
| 268-RVR | CAGCAGTCCTCAATCACCTGCTGTAC | Reverse PCR control primer |
| 281-RVR | GGCCTGTACCTCTCAGCAGTCCTC | Reverse PCR control primer |
| 293-RVR | CATAAGAATTTTGGCCTGTACC | Reverse PCR control primer |
| 363-RVR | GTGTGTGAGTTTGTGAAGAATG | Reverse PCR control primer |
| 528-RVR | CACAATTACAAATATCAGCCTGCATGTG | Reverse PCR control primer |
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Control primers anneal to a sequence near the end of theHFE (HLA-6) gene. (ASP) allele-specific primer, (WT) wild-type, (VT) variant.
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↵Indicates primer is labeled in an ET format with CYA as the donor dye covalently attached at the 5′ end.
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↵Indicates the position of the acceptor dye on the ET primer with either a ROX dye for WT primers or a R110 dye for VT primers.
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↵The HFECONFWD primer was labeled with either ROX or TAM acceptor dye.
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↵This number indicates the predicted size of the PCR product when a particular RVR primer is used.
