100 ng Firefly luciferase mRNA was added to rabbit reticulocyte lysate (RRL) (Green Hectares, Wisconsin, USA) in a reaction volume of 10 μl, as described previously (26 (link)). Blasticidin S (Sigma-Aldrich, #15205) was added simultaneously, and the in vitro translation reactions were incubated at 30°C for 30 min. Reactions were stopped by the addition of 30 μl of 1× lysis buffer from the luciferase assay kit (Promega, #E1500). The luciferase activity was detected following the manufacturer's protocol using a Synergy Neo2 plate reader (BioTek, Vermont, USA).
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