APPENDIX III Chromatographic Mobilities of Modified Nucleotides

Abstract

These figures illustrate chromatographic properties of modified nucleotides found in tRNA. The information has been compiled from the most widely used separation systems. (See Appendix IA and M. Sprinzl, F. Grüter, and D. H. Gauss. 1979. Nucleic Acids Res. 6:r1.)

Two-dimensional thin-layer chromatography was carried out on Avicel-SF cellulose thin-layer plates at 20°C. Solvent systems used were: first dimension, isobutyric acid:0.5 M ammonium hydroxide, 5:3 (v/v); second dimension, isopropanol: concentrated HCl: water, 70:15:15 (v/v/v). The relative positions of the modified nucleotides vary slightly depending on the time of development and the solvents used (freshly prepared or after repeated use). The corresponding nucleosides move faster in the first dimension and slower in the second dimension.