Table 2 Oligonucleotide primers used in the study
Sequence (5' to 3') | TA [°C] | Primer [nM] | Cycles | Product size (bp) | |
|---|---|---|---|---|---|
RT-PCR | |||||
GAPDH | Forward: TGGTCACCAGGGCTGCTT | ||||
59 | 400 | 35 | 510 | ||
Reverse: GTCTTCTGGGTGGCAGTGAT | |||||
DKK3 | Forward: AAGGCAGAAGGAGCCACGAGTGC | ||||
59 | 400 | 35 | 182 | ||
Reverse: GGCCATTTTGGTGCAGTGACCCCA | |||||
Semiquantitative realtime PCR | |||||
GAPDH | Forward: ATGGCCAGCGGGAAGAC | ||||
60 | 500 | 40 | 289 | ||
Reverse: ATGGCCAGCGGGAAGAC | |||||
DKK3 | Forward: ACAGCCACAGCCTGGTGTA | ||||
60 | 400 | 40 | 123 | ||
Reverse: CCTCCATGAAGCTGCCAAC | |||||
Methylation-specific PCR | |||||
DKK3 unmethylated | Forward: TTAGGGGTGGGTGGTGGGGT | ||||
58 | 320 | 34 | 126 | ||
Reverse: CTACATCTCCACTCTACACCCA | |||||
DKK3 methylated | Forward: GGGCGGGCGGCGGGGC | ||||
58 | 320 | 34 | 120 | ||
Reverse: ACATCTCCGCTCTACGCCCG |