Table 1 Sequence and reaction conditions of nested MSP and real-time PCR primer
From: Axin gene methylation status correlates with radiosensitivity of lung cancer cells
Name | Sequence | Length | TM | Cycle |
|---|---|---|---|---|
MSP primers | Â | Â | Â | Â |
Axin promoter (Outside primer) | F:5′GGAGGTTTTGGTTTTTTAGAGAGYGGAG 3′ | 298 bp | 55°C | 35 |
|  | R: 5′AAACCCTAACCATCCCTACCTACCRACC 3′ |  |  |  |
Axin promoter methylation | F: 5′GTAGGTTTTTGGAATGGTCGC 3′ | 144 bp | 55.7°C | 35 |
|  | R: 5′ACTAAACAAAAAACCCCGAA 3′ |  |  |  |
Axin promoter unmethylation | F: 5′ GTAGGTTTTTGGAATGGTTGTGG 3′ | 144 bp | 55.2°C | 35 |
|  | R: 5′ ACTAAACAAAAAACCCCAAA 3′ |  |  |  |
Axin intron 1 (Outside primer) | F: 5′TGTTTATAATTTTAGTTATTTGGGAAGGT 3′ | 283 bp | 55°C | 35 |
|  | R: 5′ACCCCTTATTTTTACTCACACTTCTATT 3′ |  |  | |
Axin intron 1 methylation | F:5′ GTTGAGGTAGGAGAATCG 3′ | 222 bp | 59.3°C | 35 |
|  | R:5′ TCTTCAGGAAAAATCTCG 3′ |  |  | |
Axin intron 1 unmethylation | F:5′ GTTGAGGTAGGAGAATAG 3′ | 222 bp | 59°C | 35 |
|  | R:5′ TCTTCAGGAAAAATCTAG 3′ |  |  | |
Axin intron 2 (Outside primer) | F: 5′ GGATAAATATAGAAAAGGGTTAGGAATG 3′ | 362 bp | 57.5 | 35 |
|  | R: 5′ ATAAACTAAAAAACTCCTCAAATACCAC 3′ |  |  |  |
Axin intron 2 methylation | F:5′ AAGTGAGAGTTTAGGTAGAGGAGGC3′ | 238 bp | 58°C | 35 |
|  | R:5′ CAAAAAAACTAAATACCTATAACCG 3′ |  |  |  |
Axin intron 2 unmethylation | F:5′ GAGAGTTTAGGTAGAGGAGGT 3′ | 234 bp | 59.5°C | 35 |
|  | R:5′ CAAAAAAACTAAATACCTATAACCA 3′ |  |  | |
Real-time PCR primers | Â | Â | Â | Â |
Axin primer | F: 5′- TCACCCTGGGCCAGTTCAA -3′ |  |  |  |
|  | R: 5′-CAGTCAAACTCGTCGCTCACTTTC -3′ |  |  | |
β-actin primer | F:5′- AGCACAGAGCCTCGCCTTTG -3′ |  |  |  |
|  | R:5′- ACATGCCGGAGCCGTTGT -3′ |  |  | |