PMC:3003518 / 6522-7933 JSONTXT

{"target":"http://pubannotation.org/docs/sourcedb/PMC/sourceid/3003518","sourcedb":"PMC","sourceid":"3003518","source_url":"http://www.ncbi.nlm.nih.gov/pmc/3003518","text":"Real-time polymerase chain reaction\nReal-time quantitative polymerase chain reaction (PCR) was carried out using the LightCycler PCR system (Roche Diagnostics, Meylan, France) using the DNA binding SYBR Green I dye to detect the PCR products as described previously [19]. A serial dilution was used to generate each standard curve. Each 20-μL reaction mixture contained 1× LightCycler-DNA master SYBR Green I, a specific primer, and 2 μL of cDNA. After 2 minutes of denaturation at 95°C, the MMPs, VEGF, IL-6, IL-8, cyclooxygenase-2 (COX-2), and β-actin underwent 40 reaction cycles at 95°C for 5 seconds, 55°C to 60°C for 10 seconds (annealing), and 72°C for 13 seconds. Product specificity was determined by melting curve analysis as described in the LightCycler manual. The results are calculated as ratios of gene transcripts to β-actin transcripts, with the quantity of transcripts in each sample expressed as a copy number. The ratio of MMPs, VEGF, IL-6, IL-8, and COX-2/β-actin mRNA was assigned a value of 100%, with the corresponding results calculated as relative percentages. The primers were synthesized by Bioneer Co. Ltd. (Seoul, Korea), and their sequences are listed in Table 1.\nTable 1 The sequence of polymerase chain reaction primers used in this experiment bp, base pairs; COX-2, cyclooxygenase-2; IL, interleukin; MMP, matrix metalloproteinase; VEGF, vascular endothelial growth factor.\n\n","divisions":[{"label":"Title","span":{"begin":0,"end":35}},{"label":"Table caption","span":{"begin":1195,"end":1410}}],"tracks":[{"project":"2_test","denotations":[{"id":"19883500-17697361-4203720","span":{"begin":267,"end":269},"obj":"17697361"}],"attributes":[{"subj":"19883500-17697361-4203720","pred":"source","obj":"2_test"}]}],"config":{"attribute types":[{"pred":"source","value type":"selection","values":[{"id":"2_test","color":"#93ecb9","default":true}]}]}}