Table 1.
Primer sequences for methylation-specific polymerase chain reaction
| Primer | Upstream primer 5'→3' | Downstream primer 5'→3' | T (˚C) | Length (bp) |
|---|---|---|---|---|
| MTHFR-P1 | GAGGGGTTATGAGAAAAGATTTT | ACTCCTAATCTCAATCCCAAAA | 59 | 406 |
| MTHFR-M | GTGCGGGTTTTACGTTTATC | GAAAAAACCACGTAACCGTC | 60 | 174 |
| MTHFR-U | GGGTGTGGGTTTTATGTTTATT | AAAAAAACCACATAACCATCCC | 60 | 176 |
| CBS-P1 | GTGTTAGTTTTTGTTAGTGGATATTT | ACTAAACCTAATCCCCCC | 57 | 276 |
| CBS-M | TTTTACGTGGTAGAGATCGC | AACCTACAACGAAAAACACG | 60 | 148 |
| CBS-U | TTTTATGTGGTAGAGATTGT | AACCTACAACAAAAAACACAAAC | 60 | 148 |
| MGMT-P1 | GGTATTGGGAGTTAGGATTTTA | TTTTCCTATCACAAAAATAATCC | 56 | 423 |
| MGMT-M | TATAGGTTTTGGAGGTTGTTTTTAC | TAATAAAAATCCCGATCCTACTCG | 58 | 147 |
| MGMT-U | ATAGGTTTTGGAGGTTGTTTTTATG | AATAAAAATCCCAATCCTACTCAAA | 58 | 145 |
| P16-PCR1 | TTAGATAGAAAGGTGGTATGTGG | CCAAACCTTACAAAAAAAAAAA | 55 | 319 |
| P16-M | ATTTTGAGTGAAATTTATTTATCGG | ATACAAACCCAAAACAAAACGAA | 58 | 131 |
| P16-U | ATTTTGAGTGAAATTTATTTATTGG | ATACAAACCCAAAACAAAACAAA | 58 | 131 |
| FHIT-P1 | TTTAGAAAGATTTAGAGTGGGGA | AAACTACAATTCCCAAAAAACC | 58 | 398 |
| FHIT-M | AGAAATTTAGTTAGTGGGAAGTCGT | AAAAAAATTTAAAACATAAATCGCA | 60 | 167 |
| FHIT-U | AGAAATTTAGTTAGTGGGAAGTTGT | AAAAAAATTTAAAACATAAATCACA | 60 | 167 |
| RASSF1A-M | TTAGCGTTTAAAGTTAGCGAAGTAC | AAAATCGCACCACGTATACGTA | 60 | 241 |
| RASSF1A-U | TTAGTGTTTAAAGTTAGTGAAGTATGG | CACAAAAATCACACCACATATACATA | 60 | 245 |
Abbreviations: bp: base pair; M: methylation primer; P1: first round amplification primer; T: annealing temperature; U: unmethylation primer.