Genotyping MMP Gene Variants

Nucleic acid was manually extracted from a blood sample using the DNeasy Blood and Tissue Kit (QIAGEN, Germany) according to the instruction and it`s stored at -20°C. We used nested AS-PCR to detect MMP9 rs3918242 genotypes [28 (link)]. Also, RFLP was used to detect the variations of rs1799750 of MMP1, rs11646643 and rs243864 of MMP2, rs3918253 of MMP9, rs652438 of MMP12 gene [29 (link)–31 (link)]. List of the primers, restriction enzymes and length of amplicons are shown in S1 Table. PCR reactions were performed using the AccuPower® HotStart PCR PreMix Kit (K-5050, Bioneer Corporation, Korea). The amplicons and their length were determined by agarose gel electrophoresis (C-9100-1, Bioneer Corporation, Korea) and visualized with ethidium-bromide staining (C-9036, Bioneer Corporation, Korea).