Table 2.
Primers sequences and conditions of the methylation specific PCR (MSP).
| Gene | Primers | Annealing temperature (1C) | MgCl2 | Cycles |
|---|---|---|---|---|
| CDH1 (M) | TAATTAGCGGTACGGGGGGC CGAAAACAAACGCCGAATACG | 59 | 4.5 | 32 |
| CDH1 (U) | TTAGTTAATTAGTGGTATGGGGGGTGG ACCAAACAAAAACAAACACCAAATACA | 59 | 4.5 | 32 |
| DAPK (M) | GGATAGTCGGATCGAGTTAACGTC CCCTCCCAAACGCCGA | 59 | 4.5 | 35 |
| DAPK (U) | GGAGGATAGTTGGATTGAGTTAATGTT CAAATCCCTCCCAAACACCAA | 59 | 4.5 | 35 |
| p73 (M) | GGACGTAGCGAAATCGGGGTTC ACCCCGAACATCGACGTCCG | 64 | 4.5 | 35 |
| p73 (U) | AGGGGATGTAGTGAAATTGGGGTTT ATCACAACCCCAAACATCAACATCCA | 60 | 4.5 | 35 |
| O6O6-MGMT (M) | TTTCGACGTTCGTAGGTTTTCGC GCACTCTTCCGAAAACGAAACG | 56 | 3.5 | 35 |
| O6O6-MGMT (U) | TTTGTGTTTTGATGTTTGTAGGTTTTTGT AACTCCACACTCTTCCAAAAACAAAACA | 57 | 4.5 | 35 |
| p14 (M) | GTGTTAAAGGGCGGCGTAGC AAAACCCTCACTCGCGACGA | 54 | 4.5 | 35 |
| p14 (U) | TTTTTGGTGTTAAAGGGTGGTGTAGT CACAAAAACCCTCACTCACAACAA | 56 | 4.5 | 35 |
| p15 (M) | GCGTTCGTATTTTGCGGTT CGTACAATAACCGAACGACCGA | 57 | 3.5 | 35 |
| p15 (U) | TGTGATGTGTTTGTATTTTGTGGTT CCATACAATAACCAAACAACCAA | 59 | 4.5 | 35 |
| p16 (M) | TTATTAGAGGGTGGGGCGGATCGC CCACCTAAATCGACCTCCGACCG | 68 | 1.5 | 33 |
| p16 (U) | TTATTAGAGGGTGGGGTGGATTGT CCACCTAAATCAACCTCCAACCA | 58 | 4.5 | 33 |
| FHIT (M) | TTGGGGCGCGGGTTTGGGTTTTTACGC CGTAAACGACGCCGACCCCACTA | 71–63 | 1.5 | 32 |
| FHIT (U) | TTGGGGTGTGGGTTTGGGTTTTTATG CATAAACAACACCAACCCCACTA | 64 | 1.5 | 33 |
| APC (M) | TATTGCGGAGTGCGGGTC TCAACGAACTCCCGACGA | 62 | 3.5 | 35 |
| APC (U) | GTGTTTTATTGTGGAGTGTGGGTT CCAATCAACAAACTCCCAACAA | 62 | 1.5 | 35 |
| RASSF1A (M) | TTCGTCGTTTAGTTTGGATTTTG CCGATTAAACCCGTACTTCG | 56 | 1.5 | 35 |
| RASSF1A (U) | TGTTGTTTAGTTTGGATTTTGG TACAACCCTTCCCAACACAC | 59 | 3.5 | 35 |
| RARβ (M) | TCGAGAACGCGAGCGATTCG GACCAATCCAACCGAAACGA | 62 | 1.5 | 35 |
| RARβ (U) | TTGAGAATGTGAGTGAATTGA AACCAATCCAACCAAAACAA | 59 | 1.5 | 35 |