A total of 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 by Lee et al. (2015) (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).
Free full text: Click here
