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Transcription reactions using in-house purified T7 RNA polymerase were conducted in 5 mM ATP, 2 mM UTP, 5 mM CTP, 8 mM GTP, 5 mM DTT, 40 mM Tris pH 7.9, 2.5 mM spermidine, 26 mM MgCl 2 , 0.01% (v/v) Triton X-100, 5 mM DTT, SUPERaseIn (1 U l 1

doi: 10.1007/s00394-018-1785-z 96 OttigerMNicklerMSteuerCBernasconiLHuberAChrist-CrainMet al

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First, NAD+ levels increase, which enhances mitochondrial function and energy production

The density of conjugated PEG was determined by measuring the concentration of un-reacted PEG-SH using Ellmans assay, according to a previously described method 40