Table 2.

Primer sequences

Primer name Sequence 5′-3′ Function
C282Y-WT [ii]CTGGGTGCT[iii]CCACCTGGCASP for C282Y C allele
C282Y-VT [ii]CTGGGTGCT[iii]CCACCTGGTASP for C282Y Y allele
H63D-WT [ii]TCCACACGGCGACT[iii]CTCATGASP for H63D H allele
H63D-VT [ii]TCCACACGGCGACT[iii]CTCATCASP for H63D D allele
S65C-WT [ii]AGTTCGGGGCT[iii]CCACACGGCGACTASP for S65C S allele
S65C-VT [ii]AGTTCGGGGCT[iii]CCACACGGCGACAASP for S65C C allele
HFECONFWD [ii]CGGTGGTCCGGT[iv]GACCAGAGATGCAGCGForward PCR control primer
160-RVR[v] ACATGGTTAAGGCCTGTTGCForward H63D VT primer
174-RVRTCCCCTCCTACTACACATGGForward H63D WT primer
211-RVRAGGTGTGTGGAGCCTCAACAForward S65C WT primer
201-RVRAGCCTCAACATCCTGCTCCCForward S65C VT primer
223-RVRATCATGTGACCTCTTCAGTGACCForward C282Y WT primer
232-RVRAAGGTGACACATCATGTGACCForward C282Y VT primer
244-RVRCTTGACTCAATGATTATCAGAGTGGReverse PCR control primer
268-RVRCAGCAGTCCTCAATCACCTGCTGTACReverse PCR control primer
281-RVRGGCCTGTACCTCTCAGCAGTCCTCReverse PCR control primer
293-RVRCATAAGAATTTTGGCCTGTACCReverse PCR control primer
363-RVRGTGTGTGAGTTTGTGAAGAATGReverse PCR control primer
528-RVRCACAATTACAAATATCAGCCTGCATGTGReverse PCR control primer

[i] Control primers anneal to a sequence near the end of theHFE (HLA-6) gene. (ASP) allele-specific primer, (WT) wild-type, (VT) variant.

[ii] Indicates primer is labeled in an ET format with CYA as the donor dye covalently attached at the 5′ end.

[iii] 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.

[iv] The HFECONFWD primer was labeled with either ROX or TAM acceptor dye.

[v] This number indicates the predicted size of the PCR product when a particular RVR primer is used.