|
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 |
|||||
|
TA = annealing temperature. | |||||
Veeck et al. Breast Cancer Research 2008 10:R82 doi:10.1186/bcr2151 |
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