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Each reaction contained the following components: 10 uL cell free extract (44.5 mg/ml), 25 uL 2 x buffer (containing 10 mM Mg-glutamate, 240 mM K-glutamate, 3 mM each amino acid, 100 mM HEPES, 3 mM ATP and GTP, 1.8 mM CTP and UTP, 0.4 mg/ml tRNA, 0.52 mM CoA, 0.66 NAD, 1.5 mM cAMP, 0.136 mM folinic acid, 2 mM spermidine, 60 mM 3-PGA), 2.5 uL 40 % polyethylene glycol-8000, 3 ug DNA, inducers (if required) and ddH2O up to 50 uL