Table I.
Summary of primer sequences, chromosomal locations, annealing temperatures and product sizes used for methylation-specific polymerase chain reaction analyses.
| Genes | M/U | Sequence (5′-3′) | Annealing (°C) | Product size (bp) | Ref |
|---|---|---|---|---|---|
| β-actin | – | F: TGGTGATGGAGGAGGTTTAGTAAGT | 60.0 | 133 | 20 |
| – | R: AACCAATAAAACCTACTCCTCCCTTAA | ||||
| BRCA1 | M | F: GGTTAATTTAGAGTTTCGAGAGACG | 65.0 | 182 | 21 |
| R: TCAACGAACTCACGCCGCGCAATCG | |||||
| U | F: GGTTAATTTAGAGTTTTGAGAGATG | 62.0 | 182 | ||
| R: TCAACAAACTCACACCACACAATCA | |||||
| GSTP1 | M | F: TTCGGGGTGTAGCGGTCGTC | 55.0 | 91 | 22 |
| R: GCCCCAATACTAAATCACGACG | |||||
| U | F: GATGTTTGGGGTGTAGTGGTTGTT | 55.0 | 97 | ||
| R: CCACCCCAATACTAAATCACAACA | |||||
| P16INK4A | M | F: TTATTAGAGGGTGGGGCGGATCGC | 62.0 | 150 | 23 |
| R: GACCCCGAACCGCGACCGTAA | |||||
| U | F: TTATTAGAGGGTGGGGTGGATTGT | 60.0 | 151 | ||
| R: CAACCCCAAACCACAACCATAA | |||||
| MGMT | M | F: TTTCGACGTTCGTAGGTTTTCGC | 62.0 | 81 | 24 |
| R: GCACTCTTCCGAAAACGAAACG | |||||
| U | F: TTTGTGTTTTGATGTTTGTAGGTTTTTGT | 62.0 | 93 | ||
| R: AACTCCACACTCTTCCAAAAACAAAACA | |||||
| PTEN | M | F: TTCGTTCGTCGTCGTCGTATTT | 62.0 | 207 | 25 |
| R: GCCGCTTAACTCTAAACCGCAACCG | |||||
| U | F: GTGTTGGTGGAGGTAGTTGTTT | 62.0 | 163 | ||
| R: ACCACTTAACTCTAAACCACAACCA | |||||
| RARβ2 | M | F: TCGAGAACGCGAGCGATTCG | 62.0 | 146 | 26 |
| R: GACCAATCCAACCGAAACGA | |||||
| U | F: TTGAGAATGTGAGTGATTTGA | 60.5 | 146 | ||
| R: AACCAATCCAACCAAAACAA | |||||
| CCND2 | M | F: TCGGTGTGGTTACGTTTAGC | 59.0 | 160 | 27 |
| R: TAAAACGACGCGATACAACG | |||||
| U | F: TGGTGTGGTTATGTTTAGTG | 59.0 | 150 | ||
| R: ACAATACAACATCTAAAACCAC |
M, methylated primer; U, unmethylated primer; F, forward primer; R, reverse primer; BRCA1, breast cancer 1, early onset; DNA repair associated; GSTP1, glutathione S-transferase pi 1; P16INK4A, cyclin dependent kinase inhibitor 2A; MGMT, O-6-methylguanine-DNA methyltransferase; PTEN, phosphatase and tensin homolog; RARβ2, retinoic acid receptor beta 2; CCND2, cyclin D2.