Table I.
Specific primers used in MSP and pyrosequencing, and sizes of the PCR products.
| Type of primer | Sequence | Size (bp) | Number of analyzed CpGs |
|---|---|---|---|
| MSP primer setsa | |||
| External primer set | |||
| Forward | 5′-GTGTTTTYGGGGTTTATTTGGTTGT-3′ | 186 | |
| Reverse | 5′-TACRACTCCAAAAACCCATAACTAACC-3′ | ||
| Internal methylated primer set | |||
| Forward | 5′-TGTAGTTACGTATTTATTTTTAGTGGCGTC-3′ | 112 | |
| Reverse | 5′-CGAATACGATCGAATCGAACCG-3′ | ||
| Internal unmethylated primer set | |||
| Forward | 5′-TGGTTGTAGTTATGTATTTATTTTTAGTGGTGTT-3′ | 120 | |
| Reverse | 5′-ACACCAAATACAATCAAATCAAACCAAA-3′ | ||
| Pyrosequencing primer sets | |||
| PCR primer set | |||
| Forward | 5′-GATTGGTTGTGGTCGGTAGGTGAATTTT-3′ | 235 | 19 |
| Reverse | 5′-btn-ACTCCAAAAACCCATAACTAACC-3′ | ||
| Pyrosequencing primers | |||
| Sequencing primer 1 | 5′-GTAGGTGAATTTTTAGTTAATTAG-3′ | 7 | |
| Sequencing primer 2 | 5′-GTTTGCGGAAGTTAGTTTAGATT-3′ | 11 | |
| Sequencing primer 3 | 5′-GTGTTTTCGGGGTTTATTTGGTTGT-3′ | 5 |
a
Primers published in the literature (22). MSP, methylation-specific PCR; PCR, polymerase chain reaction; btn, biotin.