Disease Markers / 2017 / Article / Tab 1 / Research Article
Role of MMP-1 (-519A/G, -1607 1G/2G), MMP-3 (Lys45Glu), MMP-7 (-181A/G), and MMP-12 (-82A/G) Variants and Plasma MMP Levels on Obesity-Related Phenotypes and Microvascular Reactivity in a Tunisian Population Table 1 Details of PCR primer sequences and RFLP conditions.
Gene Polymorphism SNP Primer sequence PCR conditions Restriction enzyme MMP-1 -519A/G rs1144393 F: 5′-CATGGTGCTATCGCAATAGGGT-3′ R: 5′-TGCTACAGGTTTCTCCACACAC-3′ Initial denaturation of 3 min at 94°C, followed by 35 cycles of 45 s at 94°C, 1 min at 60°C, 45 s at 72°C, and a final extension at 72°C for 10 min KpnI MMP-1 -1607 1G/2G rs1799750 F: 5′-TCGTGAGAATGTCTTCCCATT-3′ R: 5′-TCTTGGATTGATTTGAGATAAG TGAAATC-3′ Initial denaturation of 2 min at 95°C, followed by 35 cycles of 45 s at 95°C, 1 min at 56°C, 45 s at 72°C, and a final extension at 72°C for 10 min XmnI MMP-3 Lys45Glu (A/G) rs679620 F: 5′-CAACACCATAGCAGTAGCAGC-3′ R: 5′-CAGCCTCTCCTTCATACAGCC-3′ Initial denaturation of 3 min at 95°C, followed by 35 cycles of 1 min at 95°C, 45 s at 60°C, 45 s at 72°C, and a final extension at 72°C for 10 min TaqI MMP-7 -181A/G rs11568818 F: 5′-TGGTACCATAATGTCCTGAATG-3′ R: 5′-TCGTTATTGGCAGGAAGCACACAATGAATT-3′ Initial denaturation of 3 min at 94°C, followed by 35 cycles of 45 s at 94°C, 1 min at 56°C, 45 s at 72°C, and a final extension at 72°C for 10 min EcoRI MMP-12 -82A/G rs2276109 F: 5′-GAGATAGTCAAGGGATGATATCAGC-3′ R: 5′-AAGAGCTCCAGAAGCAGTGG-3′ Initial denaturation of 3 min at 94°C, followed by 35 cycles of 45 s at 94°C, 1 min at 60°C, 45 s at 72°C, and a final extension at 72°C for 10 min PvuII
F: forward; MMP: matrix metalloproteinase; PCR: polymerase chain reaction; R: reverse; RLFP: restriction fragment length polymorphism; SNP: single nucleotide polymorphism.