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Isoaccepting tRNA's as their mixed trialkylammonium salts have been separated electrophoretically on a normally hydrophobic support membrane. In studies on baker's yeast tRNA, the two known major Met-tRNA's have been well separated from one another and from what may be a minor Met-tRNA. An electrophoretic study of two major Met-tRNA species after their separation by reversed-phase partition chromatography indicates that the less lipophilic of the two species has the higher electrophoretic mobility. One major and four minor radioactive loci have been found on the electrophorograms of baker's yeast tRNA which had been charged with [ 14 C]phenylalanine. The nature of the Phe-tRNA's which occupy the minor loci remains to be determined. It is suggested that they may include undermethylated precursor forms of the main Phe-tRNA species of baker's yeast.