Separation of tRNA by high-performance liquid chromatography at ambient temperature

The ‘KMSKS' Motif in Tyrosyl-tRNA Synthetase Participates in the Initial Binding of tRNATyr

“…The addition of GMP to the in vitro transcription reaction results in tRNA Tyr that contains a single phosphate group on the 5‘-terminal guanosine. In vitro transcribed tRNA Tyr was purified by extraction with a phenol/chloroform/isoamyl alcohol mixture (25:24:1) as described in Avis et al (), followed by separation on a C 4 reverse-phase column (Phenomenex) using a gradient from 10 mM MgCl 2 , 10 mM sodium phosphate, pH 5.5, 1 M sodium formate (buffer A), to 10 mM sodium phosphate, pH 5.5, and 10% methanol (v/v) (buffer B) ( , ). A peak eluting at 35% buffer B was ethanol precipitated and resuspended in 10 mM MgCl 2 .…”