Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1993 Apr 25;21(8):1705–1712. doi: 10.1093/nar/21.8.1705

Synthesis and antibody-mediated detection of oligonucleotides containing multiple 2,4-dinitrophenyl reporter groups.

J Grzybowski 1, D W Will 1, R E Randall 1, C A Smith 1, T Brown 1
PMCID: PMC309404  PMID: 8493087

Abstract

A series of non-nucleoside-based 2,4-dinitrophenyl (DNP) phosphoramidites have been prepared and used in the multiple labelling of oligonucleotides during solid-phase synthesis. The length of spacer arm between the DNP label and the oligonucleotide phosphate backbone, and the number of attached DNP groups have both been varied in order to determine the optimum conditions for anti-DNP antibody binding. Detection using enzyme-linked colorimetric techniques showed sensitivity equivalent to that obtainable using biotinylated oligonucleotides.

Full text

PDF
1705

Images in this article

1708Image
on p.1708
1708Image
on p.1708
1708Image
on p.1708
1708Image
on p.1708
1709Image
on p.1709

Selected References

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

  1. Agrawal S., Christodoulou C., Gait M. J. Efficient methods for attaching non-radioactive labels to the 5' ends of synthetic oligodeoxyribonucleotides. Nucleic Acids Res. 1986 Aug 11;14(15):6227–6245. doi: 10.1093/nar/14.15.6227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Agrawal S., Zamecnik P. C. Site specific functionalization of oligonucleotides for attaching two different reporter groups. Nucleic Acids Res. 1990 Sep 25;18(18):5419–5423. doi: 10.1093/nar/18.18.5419. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cook A. F., Vuocolo E., Brakel C. L. Synthesis and hybridization of a series of biotinylated oligonucleotides. Nucleic Acids Res. 1988 May 11;16(9):4077–4095. doi: 10.1093/nar/16.9.4077. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Forster A. C., McInnes J. L., Skingle D. C., Symons R. H. Non-radioactive hybridization probes prepared by the chemical labelling of DNA and RNA with a novel reagent, photobiotin. Nucleic Acids Res. 1985 Feb 11;13(3):745–761. doi: 10.1093/nar/13.3.745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Höltke H. J., Seibl R., Burg J., Mühlegger K., Kessler C. Non-radioactive labeling and detection of nucleic acids. II. Optimization of the digoxigenin system. Biol Chem Hoppe Seyler. 1990 Oct;371(10):929–938. doi: 10.1515/bchm3.1990.371.2.929. [DOI] [PubMed] [Google Scholar]
  6. Keller G. H., Cumming C. U., Huang D. P., Manak M. M., Ting R. A chemical method for introducing haptens onto DNA probes. Anal Biochem. 1988 May 1;170(2):441–450. doi: 10.1016/0003-2697(88)90656-2. [DOI] [PubMed] [Google Scholar]
  7. Keller G. H., Huang D. P., Manak M. M. Labeling of DNA probes with a photoactivatable hapten. Anal Biochem. 1989 Mar;177(2):392–395. doi: 10.1016/0003-2697(89)90072-9. [DOI] [PubMed] [Google Scholar]
  8. Kessler C., Höltke H. J., Seibl R., Burg J., Mühlegger K. Non-radioactive labeling and detection of nucleic acids. I. A novel DNA labeling and detection system based on digoxigenin: anti-digoxigenin ELISA principle (digoxigenin system). Biol Chem Hoppe Seyler. 1990 Oct;371(10):917–927. doi: 10.1515/bchm3.1990.371.2.917. [DOI] [PubMed] [Google Scholar]
  9. Langer P. R., Waldrop A. A., Ward D. C. Enzymatic synthesis of biotin-labeled polynucleotides: novel nucleic acid affinity probes. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6633–6637. doi: 10.1073/pnas.78.11.6633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Misiura K., Durrant I., Evans M. R., Gait M. J. Biotinyl and phosphotyrosinyl phosphoramidite derivatives useful in the incorporation of multiple reporter groups on synthetic oligonucleotides. Nucleic Acids Res. 1990 Aug 11;18(15):4345–4354. doi: 10.1093/nar/18.15.4345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Mühlegger K., Huber E., von der Eltz H., Rüger R., Kessler C. Non-radioactive labeling and detection of nucleic acids. IV. Synthesis and properties of digoxigenin-modified 2'-deoxyuridine-5'-triphosphates and a photoactivatable analog of digoxigenin (photodigoxigenin). Biol Chem Hoppe Seyler. 1990 Oct;371(10):953–965. doi: 10.1515/bchm3.1990.371.2.953. [DOI] [PubMed] [Google Scholar]
  13. Nelson P. S., Sherman-Gold R., Leon R. A new and versatile reagent for incorporating multiple primary aliphatic amines into synthetic oligonucleotides. Nucleic Acids Res. 1989 Sep 25;17(18):7179–7186. doi: 10.1093/nar/17.18.7179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Roget A., Bazin H., Teoule R. Synthesis and use of labelled nucleoside phosphoramidite building blocks bearing a reporter group: biotinyl, dinitrophenyl, pyrenyl and dansyl. Nucleic Acids Res. 1989 Oct 11;17(19):7643–7651. doi: 10.1093/nar/17.19.7643. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Schubert F., Ahlert K., Cech D., Rosenthal A. One-step labelling of oligonucleotides with fluoresceine during automated synthesis. Nucleic Acids Res. 1990 Jun 11;18(11):3427–3427. doi: 10.1093/nar/18.11.3427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Seibl R., Höltke H. J., Rüger R., Meindl A., Zachau H. G., Rasshofer R., Roggendorf M., Wolf H., Arnold N., Wienberg J. Non-radioactive labeling and detection of nucleic acids. III. Applications of the digoxigenin system. Biol Chem Hoppe Seyler. 1990 Oct;371(10):939–951. doi: 10.1515/bchm3.1990.371.2.939. [DOI] [PubMed] [Google Scholar]
  17. Smith L. M., Sanders J. Z., Kaiser R. J., Hughes P., Dodd C., Connell C. R., Heiner C., Kent S. B., Hood L. E. Fluorescence detection in automated DNA sequence analysis. Nature. 1986 Jun 12;321(6071):674–679. doi: 10.1038/321674a0. [DOI] [PubMed] [Google Scholar]
  18. Urdea M. S., Warner B. D., Running J. A., Stempien M., Clyne J., Horn T. A comparison of non-radioisotopic hybridization assay methods using fluorescent, chemiluminescent and enzyme labeled synthetic oligodeoxyribonucleotide probes. Nucleic Acids Res. 1988 Jun 10;16(11):4937–4956. doi: 10.1093/nar/16.11.4937. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Vincent C., Tchen P., Cohen-Solal M., Kourilsky P. Synthesis of 8-(2-4 dinitrophenyl 2-6 aminohexyl) amino-adenosine 5' triphosphate: biological properties and potential uses. Nucleic Acids Res. 1982 Nov 11;10(21):6787–6796. doi: 10.1093/nar/10.21.6787. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Viscidi R. P., Connelly C. J., Yolken R. H. Novel chemical method for the preparation of nucleic acids for nonisotopic hybridization. J Clin Microbiol. 1986 Feb;23(2):311–317. doi: 10.1128/jcm.23.2.311-317.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Wachter L., Jablonski J. A., Ramachandran K. L. A simple and efficient procedure for the synthesis of 5'-aminoalkyl oligodeoxynucleotides. Nucleic Acids Res. 1986 Oct 24;14(20):7985–7994. doi: 10.1093/nar/14.20.7985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Will D. W., Pritchard C. E., Brown T. The synthesis of oligonucleotides that contain 2,4-dinitrophenyl reporter groups. Carbohydr Res. 1991 Sep 2;216:315–322. doi: 10.1016/0008-6215(92)84170-w. [DOI] [PubMed] [Google Scholar]

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

RESOURCES