Table 1.
PCR conditions for MMP-2, MMP-9, and TIMP-2 restriction fragment length polymorphisms.
| Gene | Primes | Restriction enzyme | |
|---|---|---|---|
| MMP-2 -1306C/T | 5′-CTTCCTAGGCTGGTCCTTACTGA-3′ (forward) | XspI | 188 + 162 + 26 bp (CT); 188 bp (CC) |
| 5′-CTGAGACCTGAAGAGCTAAAGACCT-3′ (reverse) | 162 + 26 bp (TT) | ||
| (95°C, 10 min, 35 cycles then 95°C, 45 s then 55°C, 45 s; 72°C, 45 s) | |||
| MMP-2 -790T/G | 5′-GGGTCTTTTGTGACCTCGATC-3′ (forward) | PvuI | 118 + 99 + 19 bp (TG); 118 bp (TT) |
| 5′-GGTAAAATGAGGCTGAGACCTG-3′ (reverse) | 99 + 19 bp (GG) | ||
| (95°C, 10 min, 40 cycles then 95°C, 45 s then 55°C 45 s, 72°C, 45 s) | |||
| MMP-735C/T | 5′-ATAGGGTAAACCTCCCCACATT-3′ (forward) | HinfI | 300 + 254 + 46 bp (CT); 300 bp (CC) |
| 5′-GGTAAAATGAGGCTGAGACCTG-3′ (reverse) | 254 + 46 bp (TT) | ||
| (95°C, 10 min, 40 cycles then 95°C, 45 s then 55°C 45 s, 72°C, 45 s) | |||
| TIMP-2 -418G/C | 5′-CGTCTCTTGTTGGCTGGTCA-3′ (forward) | BsoBI | 230 + 51 +23 bp (GG); 230 + 51 bp (CC) |
| 5′-CCTTCAGCTCGACTCTGGAG-3′ (reversed) | 253 + 230 + 51 + 23 bp (CC) | ||
| (94°C, 4 min, 35 cycles then 94°C, 1 min, then 56°C 45 s, 72°C, 45 s) | |||
| MMP-9 -1562C/T | 5′- GGGATAATTTGGCTCTGGTCTTCAA-3′ (forward) | SphI | 436 bp (CC); |
| 5′-CCATGGGAACCATAGAAAAGA-3′ (reversed) | 436 + 194 + 242 bp (CT) | ||
| 94°C, 10 min, 35 cycles then 95°C, 30 s, 59°C, 45 s, 72°C, 45 s | |||
MMP = matrix metalloproteinase; PCR = polymerase chain reaction; TIMP, tissue inhibitor of MMPs.