Aminoacylated tRNAs were purified by reversed-phase HPLC on an RP-18 HPLC column ( 250 mm × 10.5 mm , Merck ) equilibrated with buffer ( 20 mM ammonium acetate , pH 5.0 , 10 mM magnesium acetate , 400 mM NaCl ) using a gradient of 0 % -15 % ) ethanol.