Preparation of Protected Oligodeoxyribonucleotides Using the Phosphite-Triester Solid Phase Method of DNA Sythesis for Californium-252 Plasma Desorption Mass Spectrometry
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A solid-phase phosphite triester method for simple and fast preparation of oligodeoxyrlbonucleotides for mass spectroscopic sequencing and molecular weight is described. As the diester form normally produced is extremely volatile, the triester form must be preserved. The triester is volatile enough to be sequenced by the method pf252Cf plasma desorption mass spectroscopy (MacFarlane, R.D. and Torgerson. D.F. (1976) Science 191, 920-925). By detaching the oligomer from the solid phase under mild Conditions (4 hours ammonia, 0°C) followed by a countercurrent chloroform extraction, an acceptable yield and purity of triester is preserved for mass spectroscopy sequencing. Reverse phase HPLC was considered to be optional. The synthesis of dimethoxytrityl-thymidyl (3’-5’) thymidine was carried out as a test of this method. We are awaiting results of the MS sequencing from Catherine MacNeal at Texas A&M University.