Abstract
Deoxyribonucleoside phosphoramidites are prepared in situ from 5'-O,N-protected deoxyribonucleosides and 2-cyanoethyl N,N,N',N'-tetraisopropylphosphorodiamidite with tetrazole as catalyst, and the solutions applied directly on an automatic solid-phase DNA synthesizer. Using LCAA-CPG support and a cycle time of 12.5 min, oligonucleotides of 16-25 bases are obtained with a DMT-efficiency per cycle of 98.0-99.3%. The crude and fully deblocked products are of a purity comparable to that obtained using purified phosphoramidites. In case of d(G)16 the product was difficult to analyse and a better product was not obtained using doubly protected (O-6 diphenylcarbamoyl) guanine.
Full text
PDF












Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Barone A. D., Tang J. Y., Caruthers M. H. In situ activation of bis-dialkylaminophosphines--a new method for synthesizing deoxyoligonucleotides on polymer supports. Nucleic Acids Res. 1984 May 25;12(10):4051–4061. doi: 10.1093/nar/12.10.4051. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Caruthers M. H. Gene synthesis machines: DNA chemistry and its uses. Science. 1985 Oct 18;230(4723):281–285. doi: 10.1126/science.3863253. [DOI] [PubMed] [Google Scholar]
- Dörper T., Winnacker E. L. Improvements in the phosphoramidite procedure for the synthesis of oligodeoxyribonucleotides. Nucleic Acids Res. 1983 May 11;11(9):2575–2584. doi: 10.1093/nar/11.9.2575. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gao X., Gaffney B. L., Senior M., Riddle R. R., Jones R. A. Methylation of thymine residues during oligonucleotide synthesis. Nucleic Acids Res. 1985 Jan 25;13(2):573–584. doi: 10.1093/nar/13.2.573. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Itakura K., Rossi J. J., Wallace R. B. Synthesis and use of synthetic oligonucleotides. Annu Rev Biochem. 1984;53:323–356. doi: 10.1146/annurev.bi.53.070184.001543. [DOI] [PubMed] [Google Scholar]
- Pon R. T., Damha M. J., Ogilvie K. K. Modification of guanine bases by nucleoside phosphoramidite reagents during the solid phase synthesis of oligonucleotides. Nucleic Acids Res. 1985 Sep 25;13(18):6447–6465. doi: 10.1093/nar/13.18.6447. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sinha N. D., Biernat J., McManus J., Köster H. Polymer support oligonucleotide synthesis XVIII: use of beta-cyanoethyl-N,N-dialkylamino-/N-morpholino phosphoramidite of deoxynucleosides for the synthesis of DNA fragments simplifying deprotection and isolation of the final product. Nucleic Acids Res. 1984 Jun 11;12(11):4539–4557. doi: 10.1093/nar/12.11.4539. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Urdea M. S., Ku L., Horn T., Gee Y. G., Warner B. D. Base modification and cloning efficiency of oligodeoxyribonucleotides synthesized by the phosphoramidite method: methyl versus cyanoethyl phosphorus protection. Nucleic Acids Symp Ser. 1985;(16):257–260. [PubMed] [Google Scholar]
- Zon G., Gallo K. A., Samson C. J., Shao K. L., Summers M. F., Byrd R. A. Analytical studies of 'mixed sequence' oligodeoxyribonucleotides synthesized by competitive coupling of either methyl- or beta-cyanoethyl-N,N-diisopropylamino phosphoramidite reagents, including 2'-deoxyinosine. Nucleic Acids Res. 1985 Nov 25;13(22):8181–8196. doi: 10.1093/nar/13.22.8181. [DOI] [PMC free article] [PubMed] [Google Scholar]