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By a combination of chemical and enzymatic methods, small oligonucleotides with lengths varying from 2 to 8 nucleotides were synthesized from mononucleotides. The small oligonucleotides were then ligated with T_4 RNA ligase into six laxge ligonucleotides (9 to 19 nucleotides long) which were further ligated to form two half molecules with 35 and 41 nucleotides respectively, Finally, the two synthetic half molecules were annealed and ligated to obtain the whole molecule of yeast alanine tRNA (tRNA_y~(Ala)). Prior to this, two semi-syntheses were performed, i. e. ligation of the synthetic 5’-half molecule with the natural 3’-half molecule and that of the natural 5’-half molecule with the synthetic 3’-half molecule.
By a combination of chemical and enzymatic methods, small oligonucleotides were synthesized from mononucleotides. The small oligonucleotides were then ligated with T_4 RNA ligase into six laxge ligonucleotides (9 to 19 nucleotides long) which were further ligated to form two half molecules with 35 and 41 nucleotides respectively, Finally, the two synthetic half molecules were annealed and ligated to obtain the whole molecule of yeast alanine tRNA (tRNA_y ~ (Ala)). Prior to this, two semi-syntheses were performed, ie ligation of the synthetic 5’-half molecule with the natural 3’-half molecule and that of the natural 5’-half molecule with the synthetic 3’-half molecule.