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IRES activity verification and dual luciferase reporter assay
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IRES sequences, truncator sequences and synthetic mutant sequences were amplified and cloned and inserted into the RLUS-IRES-FLUS-pcDNA3.1(+) reporter vector. After 48 hours of transfection, the activity of luciferase was determined using a dual luciferase reporter system. For each transfected well, the firefly luciferase activity was normalized to the Renilla luciferase activity. The experiment was repeated three times independently.
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