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. Author manuscript; available in PMC: 2022 Aug 12.
Published in final edited form as: Methods Enzymol. 2021 Jul 29;658:379–406. doi: 10.1016/bs.mie.2021.06.011

Table 2.

tRNA acceptor activity assays.

RS Charging reaction S100 Charging reaction Null charging reaction
Reagent Final concentration Reagent Final concentration Reagent Final concentration
H2O to 20 μL H2O to 20 μL H2O to 20 μL
Charging Buffer 1× Charging Buffer 1× Charging Buffer 1×
DTT 1 mM DTT 1 mM DTT 1 mM
ATP 10 mM ATP 10mM ATP 10 mM
14C-Phe 19.7 μM 14C-Phe 19.7 μM 14C-Phe 19.7 μM
Phe 80 μM Phe 80 μM Phe 80 μM
Phe tRNA fraction 5 μM Phe tRNA fraction 5 μM Phe tRNA fraction 0 μM
Phe RS 1× S100 1× Phe RS 1×