TABLE I.
Primers for DNA Methylation Analyses of the Candidate Genes (M, Methylated; U, Un-Methylated Specific Primers)
| Genes and primer types | Forward (5’-3’) | Reverse (5’-3’) | Ann. tem. °C (size, Bp.) |
|---|---|---|---|
| β-Actin promoter amplification | TTGGGAGGGTAGTTTAGTTGTGGT | CAAAACAAAACACCTTTTACCCTAA | 60 (197) |
|
β-Actin, nested for 2nd round amplification in qMSP analysis |
GGTGGGTTTAGATTTAGGTTGTGTA | CTACCTACTTTTAAAAATAACAATCAC | 60 (125) |
| MB-COMT promoter amplification | GGATTTTTGAGTAAGATTAGATT | CAATATTCCACCCTAAATCTAAAA | 56 (431) |
| MB-COMT MSP M | TATTTTGGTTATCGTCGCGC | AACGAACGCAAACCGTAACG | 56a (142) |
| MB-COMT MSP U | TATTTTGGTTATTGTTGTGT | AACAAACACAAACCATAACA | 56 (142) |
| MB-COMT nested primer for sequencing | GATATTTTTAC(T)GAGGATATT or reverse of MB-COMTpromoter amplification | 56 | |
| DRD1 promoter amplification | ATTAGTTTTGGGAGTGTTTTTTTT | TCTCTCTCAAAACCCCTAAAACTTA | 56 (206) |
| DRD1 MSP M | GATTTTTTTTTTAAACGTATTTCGGC | AAACTTCGCTAAAAACAACGACTCCG | 60 (125) |
| DRD1 MSP U | GATTTTTTTTTTAAATGTATTTTGGT | AAACTTCACTAAAAACAACAACTCCA | 60 (125) |
| DRD2 Promoter Amplification | GGTTTTTGAGTTTTTAAAGGAGAAGAT | ACACTAAAACTAAACAACTCTA | 56 (304) |
| DRD2 MSP M | CGTTTAGGTCGGGGATCGTCG | GACGCCCGAACGCGAAAAACGCG | 67 (118) |
| DRD2 MSP U | TGTTTAGGTTGGGGATTGTTG | AACACCCAAACACAAAAAACACA | 56 (118) |
| DRD2, nested for sequencing | GGATTTAGTTTGTAATTATAGT | – | 60 |
| DRD3 M | GTTAAATAAGAGTTAGTTTTTTTATAATTGTATC | TCCTCTATTTAATCAATATAATAATTCTTCG | 60 (105) |
| DRD3 U | GTTAAATAAGAGTTAGTTTTTTTATAATTGTATT | TCCTCTATTTAATCAATATAATAATTCTTCA | 60 (105) |
| DRD4 promoter amplification | GGTAGAGTT TGAGTTTAGGTT TTTT | AAACCCAATATTTACTCATCTTAA | 56 (341) |
| DRD4 M | CGTTCGAGGGTCGGGAC | GAACAAACCAACATCGCCCGACG | 60 (151) |
| DRD4 U | TGTTTGAGGGTTGGGAT | AAACAAACCAACATCACCCAACA | 60 (151) |
| AKT1 M | GTAGAGCGAGTTTAGAAGCGTC | ACTAAACCGCTAACGAAACCCG | 60 (112) |
| AKT1 U | GTAGAGTGAGTTTAGAAGTGTT | ACTAAACCACTAACAAAACCCA | 60 (112) |
a
Annealing temperature for qMSP at 62°C.