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. 1977 Apr;4(4):1135–1158. doi: 10.1093/nar/4.4.1135

Rapid synthesis of oligodeoxyribonucleotides: a new solid-phase method.

M J Gait, R C Sheppard
PMCID: PMC342510  PMID: 866189

Abstract

A method is described that makes use of a new polyamide resin for the rapid synthesis of short oligodeoxyribonucleotides. The method is illustrated by the preparation of two heptadeoxyribonucleotides, d(pT6-C) and d(pC-A-G-T-G-A-T) using a phosphodiester approach. A further development involved use of phenyl isocyanate as an in situ drying agent, which obviated the need for solvent co-evaporation prior tothe internucleotidic coupling steps. Improved fractionation of thymidyl oligonucleotides was obtained by use of a new microparticulate, silica-based anion-exchanger.

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Selected References

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  1. Agarwal K. L., Berlin Y. A., Fritz H. J., Gait M. J., Kleid D. G., Lees R. G., Norris K. E., Ramamoorthy B., Khorana H. G. Studies on polynucleotides. CXLIII. A rapid and convenient method for the synthesis of deoxyribooligonucleotides carrying 5'-phosphate end groups using a new protecting group. J Am Chem Soc. 1976 Mar 3;98(5):1065–1072. doi: 10.1021/ja00421a001. [DOI] [PubMed] [Google Scholar]
  2. Agarwal K. L., Khorana H. G. Studies on polynucleotides. CII. The use of aromatic isocyanates for selective blocking of the terminal 3'-hydroxyl group in protected deoxyribooligonucleotides. J Am Chem Soc. 1972 May 17;94(10):3578–3585. doi: 10.1021/ja00765a054. [DOI] [PubMed] [Google Scholar]
  3. Arentzen R., Reese C. B. The phosphotriester approach to oligonucleotide synthesis: Preparation of oligo- and poly-thymidylic acids. J Chem Soc Perkin 1. 1977;4:445–460. doi: 10.1039/p19770000445. [DOI] [PubMed] [Google Scholar]
  4. Asteriadis G. T., Armbruster M. A., Gilham P. T. Separation of oligonucleotides, nucleotides, and nucleosides on columns of polystyrene anion-exchangers with solvent systems containing ethanol. Anal Biochem. 1976 Jan;70(1):64–74. doi: 10.1016/s0003-2697(76)80048-6. [DOI] [PubMed] [Google Scholar]
  5. Atherton E., Bridgen J., Sheppard R. C. A polyamide support for solid-phase protein sequencing. FEBS Lett. 1976 Apr 15;64(1):173–175. doi: 10.1016/0014-5793(76)80276-1. [DOI] [PubMed] [Google Scholar]
  6. Atherton E., Clive D. L., Sheppard R. C. Letter: Polyamide supports for polypeptide synthesis. J Am Chem Soc. 1975 Oct 29;97(22):6584–6585. doi: 10.1021/ja00855a053. [DOI] [PubMed] [Google Scholar]
  7. Blackburn G. M., Brown M. J., Harris M. R. Synthetic studies of nucleic acids on polymer supports. I. Oligodeoxyribonucleotide synthesis on an insoluble polymer support. J Chem Soc Perkin 1. 1967;22:2438–2442. doi: 10.1039/j39670002438. [DOI] [PubMed] [Google Scholar]
  8. Brownlee G. G., Sanger F. Chromatography of 32P-labelled oligonucleotides on thin layers of DEAE-cellulose. Eur J Biochem. 1969 Dec;11(2):395–399. doi: 10.1111/j.1432-1033.1969.tb00786.x. [DOI] [PubMed] [Google Scholar]
  9. Büchi H., Khorana H. G. CV. Total synthesis of the structural gene for an alanine transfer ribonucleic acid from yeast. Chemical synthesis of an icosadeoxyribonucleotide corresponding to the nucleotide sequence 31 to 50. J Mol Biol. 1972 Dec 28;72(2):251–288. doi: 10.1016/0022-2836(72)90148-9. [DOI] [PubMed] [Google Scholar]
  10. Egan B. Z. Separation of oligonucleotides by reversed-phase chromatography. Biochim Biophys Acta. 1973 Mar 19;299(2):245–252. doi: 10.1016/0005-2787(73)90347-x. [DOI] [PubMed] [Google Scholar]
  11. Greene P. H., Poonian M. S., Nussbaum A. L., Tobias L., Garfin D. E., Boyer H. W., Goodman H. M. Restriction and modification of a self-complementary octanucleotide containing the EcoRI substrate. J Mol Biol. 1975 Dec 5;99(2):237–261. doi: 10.1016/s0022-2836(75)80143-4. [DOI] [PubMed] [Google Scholar]
  12. Hartwick R. A., Brown P. R. The performance of microparticle chemically-bonded anion-exchange resins in the analysis of nucleotides. J Chromatogr. 1975 Oct 29;112:650–662. doi: 10.1016/s0021-9673(00)99994-1. [DOI] [PubMed] [Google Scholar]
  13. Hayatsu H., Khorana H. G. Studies on polynucleotides. LXXII. Deoxyribooligonucleotide synthesis on a polymer support. J Am Chem Soc. 1967 Jul 19;89(15):3880–3887. doi: 10.1021/ja00991a035. [DOI] [PubMed] [Google Scholar]
  14. Itakura K., Katagiri N., Bahl C. P., Wightman R. H., Narang S. A. Improved triester approach for the synthesis of pentadecathymidylic acid. J Am Chem Soc. 1975 Dec 10;97(25):7327–7332. doi: 10.1021/ja00858a020. [DOI] [PubMed] [Google Scholar]
  15. Kaiser E., Colescott R. L., Bossinger C. D., Cook P. I. Color test for detection of free terminal amino groups in the solid-phase synthesis of peptides. Anal Biochem. 1970 Apr;34(2):595–598. doi: 10.1016/0003-2697(70)90146-6. [DOI] [PubMed] [Google Scholar]
  16. Khorana H. G., Agarwal K. L., Besmer P., Büchi H., Caruthers M. H., Cashion P. J., Fridkin M., Jay E., Kleppe K., Kleppe R. Total synthesis of the structural gene for the precursor of a tyrosine suppressor transfer RNA from Escherichia coli. 1. General introduction. J Biol Chem. 1976 Feb 10;251(3):565–570. [PubMed] [Google Scholar]
  17. Knorre D. G., Zarytova V. F. The mechanism of the chemical synthesis of oligonucleotides and its synthetic consequences. Nucleic Acids Res. 1976 Oct;3(10):2709–2729. doi: 10.1093/nar/3.10.2709. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Letsinger R. L., Mahadevan V. Stepwise synthesis of oligodeoxyribonucleotides on an insoluble polymer support. J Am Chem Soc. 1966 Nov 20;88(22):5319–5324. doi: 10.1021/ja00974a053. [DOI] [PubMed] [Google Scholar]
  19. Marians K. J., Wu R., Stawinski J., Hozumi T., Narang S. A. Cloned synthetic lac operator DNA is biologically active. Nature. 1976 Oct 28;263(5580):744–748. doi: 10.1038/263744a0. [DOI] [PubMed] [Google Scholar]
  20. Melby L. R., Strobach D. R. Oligonucleotide synthesis on insoluble polymer supports. I. Stepwise synthesis of trithymidine diphosphate. J Am Chem Soc. 1967 Jan 18;89(2):450–453. doi: 10.1021/ja00978a052. [DOI] [PubMed] [Google Scholar]
  21. Narang S. A., Bhanot O. S., Goodchild J., Wightman R. H., Dheer S. K. A new general method for the synthesis of phosphate-protected deoxyribooligonucleotides. IV. J Am Chem Soc. 1972 Aug 23;94(17):6183–6189. doi: 10.1021/ja00772a042. [DOI] [PubMed] [Google Scholar]
  22. Pless R. C., Letsinger R. L. Solid support synthesis of oligothymidylates using phosphorochloridates and 1-alkylimidazoles. Nucleic Acids Res. 1975 Jun;2(6):773–786. doi: 10.1093/nar/2.6.773. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. TOMLINSON R. V., TENER G. M. THE EFFECT OF UREA, FORMAMIDE, AND GLYCOLS ON THE SECONDARY BINDING FORCES IN THE ION-EXCHANGE CHROMATOGRAPHY OF POLYNUCLEOTIDES OF DEAE-CELLULOSE. Biochemistry. 1963 Jul-Aug;2:697–702. doi: 10.1021/bi00904a013. [DOI] [PubMed] [Google Scholar]

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