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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

Compared with patients without lung involvement, the occurrence of lung metastasis significantly reduces the survival of patients and increases the risk of death

Kumar, D.M., Agarwal