Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1986 Oct 24;14(20):7985–7994. doi: 10.1093/nar/14.20.7985

A simple and efficient procedure for the synthesis of 5'-aminoalkyl oligodeoxynucleotides.

L Wachter, J A Jablonski, K L Ramachandran
PMCID: PMC311829  PMID: 3774550

Abstract

Synthetic deoxyoligonucleotides have been 5'-aminoalkylated at the end of step-wise synthesis on the polymer support. This was achieved through the activation of the 5'-hydroxyl group as its 5'-imidazolyl derivative using carbonyldiimidazole, which was subsequently displaced with hexamethylene diamine to yield the title compound. The alkyl carbamate linkage thus generated withstands the deprotection conditions used in oligonucleotide synthesis. Purification by gel electrophoresis and further derivatization at the 5'-amino group with N-hydroxysuccinimidobiotin is described.

Full text

PDF

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bethell G. S., Ayers J. S., Hancock W. S., Hearn M. T. A novel method of activation of cross-linked agaroses with 1,1'-carbonyldiimidazole which gives a matrix for affinity chromatography devoid of additional charged groups. J Biol Chem. 1979 Apr 25;254(8):2572–2574. [PubMed] [Google Scholar]
  2. Chollet A., Kawashima E. H. Biotin-labeled synthetic oligodeoxyribonucleotides: chemical synthesis and uses as hybridization probes. Nucleic Acids Res. 1985 Mar 11;13(5):1529–1541. doi: 10.1093/nar/13.5.1529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chu B. C., Orgel L. E. Detection of specific DNA sequences with short biotin-labeled probes. DNA. 1985 Aug;4(4):327–331. doi: 10.1089/dna.1985.4.327. [DOI] [PubMed] [Google Scholar]
  4. Connolly B. A., Rider P. Chemical synthesis of oligonucleotides containing a free sulphydryl group and subsequent attachment of thiol specific probes. Nucleic Acids Res. 1985 Jun 25;13(12):4485–4502. doi: 10.1093/nar/13.12.4485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fahrenholtz K. E., Boris A., Kennedy T. W., Jr, Kierstead R. W. Steroidal imidazole-1-carboxylic acid esters. J Med Chem. 1974 Mar;17(3):337–342. doi: 10.1021/jm00249a018. [DOI] [PubMed] [Google Scholar]
  6. Hearn M. T., Bethell G. S., Ayers J. S., Hancock W. S. Application of 1,1'-carbonyldiimidazole-activated agarose for the purification of proteins. II. The use of an activated matrix devoid of additional charged groups for the purification of thyroid proteins. J Chromatogr. 1979 Dec 20;185:463–470. doi: 10.1016/s0021-9673(00)85622-8. [DOI] [PubMed] [Google Scholar]
  7. Hearn M. T., Harris E. L., Bethell G. S., Hancock W. S., Ayers J. A. Application of 1,1'-carbonyldiimidazole-activated matrices for the purification of proteins. III. The use of 1,1'-carbonyldiimidazole-activated agaroses in the biospecific affinity chromatographic isolation of serum antibodies. J Chromatogr. 1981 Nov 20;218:509–518. doi: 10.1016/s0021-9673(00)82076-2. [DOI] [PubMed] [Google Scholar]
  8. Kempe T., Sundquist W. I., Chow F., Hu S. L. Chemical and enzymatic biotin-labeling of oligodeoxyribonucleotides. Nucleic Acids Res. 1985 Jan 11;13(1):45–57. doi: 10.1093/nar/13.1.45. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Leary J. J., Brigati D. J., Ward D. C. Rapid and sensitive colorimetric method for visualizing biotin-labeled DNA probes hybridized to DNA or RNA immobilized on nitrocellulose: Bio-blots. Proc Natl Acad Sci U S A. 1983 Jul;80(13):4045–4049. doi: 10.1073/pnas.80.13.4045. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Smith L. M., Fung S., Hunkapiller M. W., Hunkapiller T. J., Hood L. E. The synthesis of oligonucleotides containing an aliphatic amino group at the 5' terminus: synthesis of fluorescent DNA primers for use in DNA sequence analysis. Nucleic Acids Res. 1985 Apr 11;13(7):2399–2412. doi: 10.1093/nar/13.7.2399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Sproat B. S., Brown D. M. A new linkage for solid phase synthesis of oligodeoxyribonucleotides. Nucleic Acids Res. 1985 Apr 25;13(8):2979–2987. doi: 10.1093/nar/13.8.2979. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES