5 μl in vitro translation reactions were set up with 2.5 μl of lysate and 20 ng total RNA (0.84 mM ATP, 0.21 mM GTP, 21 mM creatine phosphate, 0.009 units/ml creatine phosphokinase, 10 mM HEPES pH 7.5, 2 mM DTT, 2 mM MgOAc, 100 mM KOAc, 0.008 mM amino acids, 0.25 mM spermidine, 5 units RNasin Plus RNase inhibitor (Promega) as described (48 (link)). Reaction tubes were incubated at 30°C for 45 min, and expression of the reporter was measured using the Renilla Luciferase Assay System (Promega) on a GloMax Explorer plate reader (Promega).
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