Synthesis of oligonucleotides on cellulose by a phosphotriester method.
The synthesis of oligothymidilic acids, (dT)m (where m = 4, 7, 10, 13, 16, 19, 22, and 25), was carried out using a solid phase approach in combination with the modified phosphotriester methodology developed in solution. Cellulose was used as the solid support after its functionalization with a specially featured dinucleoside diphosphate, 5'-0-p-chlorophenylphospho-2'(3')-0-acetyluridilyl-[2'(3')-3']-5'-0-dimethoxytritylthymidine p-chlorophenylester. The fully protected trideoxynucleoside triphosphate containing only thymidine was repeatedly used to elongate the oligonucleotide chain in the 3'-5' direction. Individual coupling yields ranged from 45% to 75%. The total time needed to prepare (dT)25 was four days. Similarly, the tridecanucleotide d(AGAAGGTACTTTT) was synthesized in good yield. The results show that this approach can be used for a fast and economic way to synthesize oligodeoxynucleotides.
D002482:Cellulose; D008722:Methods; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis*; D009841:Oligonucleotides / Q000138:chemical synthesis*; D010755:Organophosphates; D011071:Poly T / Q000138:chemical synthesis*; D011089:Polydeoxyribonucleotides / Q000138:chemical synthesis*
D009838:Oligodeoxyribonucleotides; D009841:Oligonucleotides; D010755:Organophosphates; D011089:Polydeoxyribonucleotides; D011071:Poly T; D002482:Cellulose