Table I.
Primers and TaqMan allele-specific probes
| Polymorphism | 5’-3’ sequence |
|---|---|
| RFC-1 80G>A | |
| Forward | GGCCTGACCCCG AGCT |
| Reverse | AGCCGTAGAAGCAAAGGTAGCA |
| G-Probe 1 | VIC-CACGAG GCGCCGC |
| A-Probe 2 | 6FAM-CGAGGT GCCGCCAG |
| MTHFR C677T | |
| Forward | TGGCAGGTTACCCCAAAGG |
| Reverse | CACAAAGCGGAAGAATGTGTCA |
| T- Probe 1 | 6FAM-TGATGAAATCGGCTCCCGCA-TAMRA |
| C- Probe 2 | VIC-TGA TGATGAAATCGACTCCCGCA-TAMRA |
| MTHFR A1298C | |
| Forward | GGAGGAGCTGCTGAAGATGTG |
| Reverse | CCCGAGAGGTAAAGAACAAAGACTT |
| A- Probe 1 | VIC-ACCAGTGAAGAAAGTGT |
| C-Probe 2 | 6FAM-CAGTGAAGCAAGTGT |
| MTRR A66G | |
| Forward | AGCAGGGACAGGCAAAGG |
| Reverse | AAGATCTGCAGAAAATCCATGTACCA |
| A-Probe 1 | VIC-TTGCTCACATATTTCTT |
| G-Probe 2 | 6FAM-CTCACACATTTCTT |
| TCN2 C776G | |
| Forward | ACTCTATCACCAGTTCCTCATGACT |
| Reverse | TTGAGACATGCTGTTCCCAGTT |
| C- Probe 1 | VIC- CTGCCCCAGGCATG |
| G-Probe 2 | 6FAM- CTGCCCCACGCATG |
Allelic discrimination was accomplished when fluorogenic probes with either a 6FAM or VIC reporter dye attached to the 5’ end of the oligonucleotide was cleaved by the 5’ nuclease activity of Taq DNA polymerase.