Table 1 Primer sequences and PCR conditions for bisulphite sequencing (SEQ), methylation- (MSP) and unmethylation-specific PCR (USP) and RT–PCR for SOX9
Sense primer (5′ → 3′) | Antisense primer (5′ → 3′) | Product size (bp) | Annealing temperature (PCR cycles) | |
|---|---|---|---|---|
SEQ | GGGGTTGGAGAATGATTTGTTAGAG | TCTAAACTAAAATCTACCCCCACCTC | 434 | 62 (40) |
MSP | GGTAGGTAGGTTCGTTTTAGGC | TCTCTCCGCGACACCGAAAA | 152 | 58 (34) |
USP | GTAGGTAGGTAGGTTTGTTTTAGGT | CTCTCTCCACAACACCAAAAACT | 152 | 58 (34) |
RT–PCR | AGTACCCGCACTTGCACAAC | CGTTCTTCACCGACTTCCTC | 178 | 60 (25) |
