Table 2. Primer sequences and PCR conditions.
| Sense primer (5'-3') | Antisense primer (5'-3') | Anneal temperature (°C), PCR cycles | Product size (bp) | |
|---|---|---|---|---|
| MSP primers | ||||
| Univiersal | ||||
| Wif-1 PAN | GTTAGTTTTGTTAGTTTTA | CTAAATACCAAAAAACCTAC | 48, 45 | 149 |
| Methylated | ||||
| Wif-1 MSP | TTTTGTCGTTTTTATTTTCGTTC | AAAAAAACAAACAAAACGCG | 52, 45 | 96 |
| Un-methylated | ||||
| Wif-1 USP | GTTTTTGTTGTTTTTATTTTTGTTT | AAAAAAAACAAACAAAACACAA | 52, 45 | 99 |
| RT-PCR primer | ||||
| β-catenin (mutation analysis) | AAGCGGCTGTTAGTCACTGG | CAGGACTTGGGAGGTATCCA | 52, 45 | 134 |
Note: Boldface indicates the differences between modified sequence. Underline indicates the changes between methylated and un-methylated sequences after bisulfite modification.