PMC:4536246 / 9398-10328 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/4536246","sourcedb":"PMC","sourceid":"4536246","source_url":"https://www.ncbi.nlm.nih.gov/pmc/4536246","text":"DDAH-1 mRNA expression was analyzed by a real-time polymerase chain reaction (RT-PCR) and total RNA was isolated from the cerebral samples with Trizol reagent in accordance with the method of Chomczynski and Mackey [26]. RNA was quantified by measuring the absorbance at 260/280 nm. cDNA was generated using the iScript cDNA Synthesis kit (Bio-Rad) following the supplier's instructions. Gene expression was analyzed using the Fast Eva Green supermix (Bio-Rad). As regards housekeeping, gene glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used. The expression of the housekeeping gene remained constant in all the experimental groups considered. The amplification was performed through two-step cycling (95–60 °C) for 45 cycles in a light Cycler 480 Instrument RT-PCR Detection System (Roche) following the supplier's instructions. All samples were assayed in triplicate. Gene expression was calculated using the ΔCt method.","tracks":[{"project":"TEST0","denotations":[{"id":"26272684-216-222-1511556","span":{"begin":216,"end":218},"obj":"[\"7539982\"]"}],"attributes":[{"subj":"26272684-216-222-1511556","pred":"source","obj":"TEST0"}]},{"project":"2_test","denotations":[{"id":"26272684-7539982-60565584","span":{"begin":216,"end":218},"obj":"7539982"}],"attributes":[{"subj":"26272684-7539982-60565584","pred":"source","obj":"2_test"}]}],"config":{"attribute types":[{"pred":"source","value type":"selection","values":[{"id":"TEST0","color":"#ecae93","default":true},{"id":"2_test","color":"#9493ec"}]}]}}