Table II.
Methylation-Specific PCR (MSP) Primers
| Primer Designation | Human/Mouse | Primer Sequence (5′ ➝ 3′) | Orientation (Forw/Rev) | Annealing Temp (°C) | Thermo- cycles | Product Size (bp) |
|---|---|---|---|---|---|---|
| Timp3-MSP-M | M | CGGAGGGGGCGGATTTC | F | 60 | 40 | 84 |
| CACGAATCCGCGAAACCAAC | R | |||||
| Timp3-MSP-U | M | TGGAGGGGGTGGATTTT | F | 55 | 40 | 184 |
| ACACAAATCCACAAAACCAAC | R | |||||
| Rprm-MSP-M | M | GTCGGGTATTTGTTCGTTTCGC | F | 58 | 40 | 96 |
| ACTAAATTCATCACGAAAACAAACGAAAACCG | R | |||||
| Rprm-MSP-U | M | GTTAGTGTTTTGTTGGGTATTTGTTTGTTTTGTG | F | 58 | 40 | 96 |
| ACACTAAATTCATCACAAAAACAAACAAAAACCA | R | |||||
| Smpd3-MSP-M | M | GGGTTTTTTAGAGACGGTTTTCGC | F | 59 | 40 | 90 |
| GCTCCAAATACTAAATCCGAAAAAAACTCCG | R | |||||
| Smpd3-MSP-U | M | TGATTGGGGTTTTTTAGAGATGGTTTTTGTG | F | 58 | 40 | 90 |
| CCACTCCAAATACTAAATCCAAAAAAAACTCCA | R | |||||
| Atp1b2-MSP-M | M | CGTGGAGGGTTTTAGCGC | F | 58 | 40 | 74 |
| AAACACGAAACGAATCCCGACG | R | |||||
| Aatp1b2-MSP-U | M | GTGTGTGGAGGGTTTTAGTGTG | F | 57.5 | 40 | 74 |
| AACCATCTTAAAAACACAAAACAAATCCCAACA | R | |||||
| Dusp2-MSP-M | M | GTTCGTTTTTGGTTTTTTGTCGGAGTTAC | F | 58.5 | 40 | 102 |
| AAAACAACGACGAAAATACCGCG | R | |||||
| Dusp2-MSP-U | M | TGTTGTTTGTTTTTGGTTTTTTGTTGGAGTTATG | F | 58 | 40 | 102 |
| ACAAACAAACAAAAACAACAACAAAAATACCACA | R | |||||
| FoxJ1-MSP-M | M | GGTTTTTTTAGCGGTTAAATTCGACGC | F | 59 | 40 | 86 |
| GCTAAATCTCACGACGAATACTCG | R | |||||
| FoxJ1-MSP-U | M | GGTTGGGTTTTTTTAGTGGTTAAATTTGATGTG | F | 58 | 40 | 86 |
| CCCACACTAAATCTCACAACAAATACTCA | R | |||||
| SMPD3-MSP-M | H | TTAGATTTGCGGTCGGGAGTTC | F | 58.5 | 40 | 100 |
| AACACTAACCTCTAACGACGAACG | R | |||||
| SMPD3-MSP-U | H | TTGTTGTTAGATTTGTGGTTGGGAGTTTG | F | 58.5 | 40 | 100 |
| AAATTCAACAACACTAACCTCTAACAACAAACA | R | |||||
| ATP1B2-MSP-M | H | GGCGATCGCGGGTTTC | F | 59 | 40 | 116 |
| CCGATAATAACTCAACCGCGAAAATAAAACG | R | |||||
| ATP1B2-MSP-U | H | GTATAAAGATTGTGTTTGGTGATTGTGGGTTTTG | F | 58.5 | 40 | 116 |
| ACACCAATAATAACTCAACCACAAAAATAAAACA | R | |||||
| DUSP2-MSP-M | H | TTATCGTTACGAGAGTTCGGGAC | F | 58 | 40 | 119 |
| CGCAACAACGTACCCAACG | R | |||||
| DUSP2-MSP-U | H | GGTAGTGTTATTGTTATGAGAGTTTGGGATG | F | 58.5 | 40 | 119 |
| AAATCCCACAACAACATACCCAACA | R | |||||
| FOXJ1-MSP-M | H | TTTTTAGAACGAACGGATTTATAGC | F | 59 | 40 | 193 |
| ACACTTACCACTCTCCCTAACGAC | R | |||||
| FOXJ1-MSP-U | H | TTTAGAATGAATGGATTTATAGTGG | F | 58 | 40 | 193 |
| AACACTTACCACTCTCCCTAACAAC | R | |||||
| Ecad-MSP-M | M | GGTTTAATTCGGTTTTGTTCGATCGTATTC | F | 58 | 40 | 75 |
| GAACTCCCATAACGAACCCG | R | |||||
| Ecad-MSP-U | M | GTTTGGTTTAATTTGGTTTTGTTTGATTGTATTTG | F | 58 | 40 | 75 |
| ACCAAACTCCCATAACAAACCCA | R | |||||
| RassF1-MSP-M | M | GGGCGGTCGTTGGAATTTC | F | 58.5 | 40 | 99 |
| CCCTCCGAATAACTCGCTATAAAAAAACG | R | |||||
| RassF1-MSP-U | M | AGGGGTGGTTGTTGGAATTTTG | F | 57.5 | 40 | 99 |
| CAAAACCCTCCAAATAACTCACTATAAAAAAACA | R | |||||
| Gstp1-MSP-M | M | GAATTCGTATTTAGTAGGTAGTGAGTTCGTTTC | F | 58 | 40 | 106 |
| CGATACAAAAAACGCCAATCGTAAACG | R | |||||
| Gstp1-MSP-U | M | TGAATTTGTATTTAGTAGGTAGTGAGTTTGTTTTG | F | 58 | 40 | 106 |
| AACTACCCAATACAAAAAACACCAATCATAAACA | R | |||||
| Rarβ-MSP-M | M | GATTTTAAGTTTTTTTTTTTAGGCGACGTTC | F | 56.5 | 40 | 89 |
| GAACAAAAAACGTCTAAACACCG | R | |||||
| Rarβ-MSP-U | M | ATTGATTTTAAGTTTTTTTTTTTAGGTGATGTTTG | F | 56.5 | 40 | 89 |
| CCCAAACAAAAAACATCTAAACACCA | R | |||||
| Dapk1-MSP-M | M | TTCGGGTCGCGGAGTC | F | 59 | 40 | 110 |
| CAATTCCTCGCCGATATCGTAATAATCG | R | |||||
| Dapk1-MSP-U | M | GATGGTTTGGGTTGTGGAGTTG | F | 57.5 | 40 | 110 |
| CTACCCAATTCCTCACCAATATCATAATAATCA | R | |||||
| Wif1-MSP-M | M | GGGTATCGCGGTTTCGTTTTC | F | 58.5 | 40 | 96 |
| CTACTAACAAAAAACGAACACTAAAAACGACCG | R | |||||
| Wif1-MSP-U | M | GGGTTTATTGGGTATTGTGGTTTTGTTTTTG | F | 58 | 40 | 96 |
| ACTACTAACAAAAAACAAACACTAAAAACAACCA | R |