Nonradioactive Labeling of Polymerase Chain Reaction Products

  • Udo Reischl
  • Rüdiger Rüger
  • Christoph Kessler
Part of the Methods in Molecular Biology book series (MIMB, volume 15)


Polymerase chain reaction (PCR) was originally introduced to amplify in vitro particular DNA sequences by the application of temperature cycles (1). In a modification, RNA molecules also may serve as templates by an additional reverse transcription step converting RNA in complementary DNA sequences (2).


Maleate Buffer Polymerase Chain Reaction Thermal Cycler Label Polymerase Chain Reaction Product Efficient Polymerase Chain Reaction Amplification Renaturation Solution 
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  1. 1.
    Saiki, R. K., Scharf, S., Faloona, F., Mullis, K. B., Horn, G. T., Erlich, H. A., and Amheim, N. (1985) Enzymatic amplification of ß-globin sequences and restrrction site analysis for diagnosis of sickle cell anemia. Scrence 230, 1350–1354.CrossRefGoogle Scholar
  2. 2.
    Murakawa, G. J., Wallace, B. R., Zaia, J. A., and Rossi, J. J. (1987) Method for amplification and detection of RNA sequences. European Patent Application 0272098Google Scholar
  3. 3.
    Wu, D. Y. and Wallace, R. B, (1989) The ligation and amplification reaction (LAR)—amplification of specific DNA sequences using sequential rounds of template dependent ligation. Genomics 4, 560–569.PubMedCrossRefGoogle Scholar
  4. 4.
    Segev, D. (1990) Amplification and detection of target nucleic acid sequences— for in vitro diagnosis of infectious disease, genetic disorders or cellular disorders, e.g., cancer. Published under Patent Corporation Treaty (PCT) International Application WO 90/01069.Google Scholar
  5. 5.
    Kwoh, D. Y., Davis, G. R., Whitfield, K. M., Chapelle, H. L., DiMichelle, L. J., and Gingeras, T. R. (1989) Transcription-based amplification system and detection of amplified human immunodeficiency virus. Proc. N&l. Acad, Sci. USA 86, 1173–1177.CrossRefGoogle Scholar
  6. 6.
    Guatelli, J. C., Whitfield, K. M., Kwoh, D. Y., Barringer, K. J., Richman, D. D., and Gingeras, T. R. (1990) Isothermal, in vitro amplification of nucleic acids by a multienzyme reaction modeled after retroviral replication. Proc. Natl. Acad. Sci. USA 87, 1874–1878.PubMedCrossRefGoogle Scholar
  7. 7.
    Davey, C. and Malek, L. T. (1988) Nucleic acid amplification process. European Patent Application 0329098.Google Scholar
  8. 8.
    Schuster, D., Thornton, C., Buchmann, G., Berninger, M., and Rashtchian, A. (1990) A method for isothermal amplification of nucleic acid sequences, 5th San Diego Conference on Nucleic Acids, American Association of Clinical Chemistry (AACC), Abstract Poster 40.Google Scholar
  9. 9.
    Lizardi, P. M., Guerra, C. E., Lomeli, H., Tussie-Luna, I., and Kramer, F. R. (1988) Exponential amplification of recombinant RNA hybridization probes. Biotechnology 6, 1197–1202.CrossRefGoogle Scholar
  10. 10.
    Saiki, R. K., Gelfand, D. H., Stoffel, S., Scharf, S. H., Higuchi, R., Horn, G. T., Mullis, K. B., and Erlich, H.A.(1988) Primer-directed enzymatic ampliflcation of DNA with a thermostable DNA polymerase. Science 239, 487–491.PubMedCrossRefGoogle Scholar
  11. 11.
    Kessler, C. (1990) Detection of nucleic acids by enzyme-linked immuno-sorbent assay (ELISA) technique: An example for the development of a novel non-radioactive labeling and detection system with high sensitivity, in Advances in Mutagenesls Research (Obe, G., ed.), Springer-Verlag, Berlin, pp. 105–152.CrossRefGoogle Scholar
  12. 12.
    Kessler, C, Hòltke, H.-J., Seibl, R., Burg, J., and Muhlegger, K. (1990) 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 371, 917–927.PubMedCrossRefGoogle Scholar
  13. 13.
    Hòltke, H.-J., Seibl. R., Burg, J., Mhhlegger, K., and Kessler, C. (1990) Non-radioactive labeling and detection of nucleic acids: II. Optimization of the digoxigenin system. Blol. Chem. Hoppe-Seyler 371, 929–938.CrossRefGoogle Scholar
  14. 14.
    Seibl. R., Höltke, H.-J., Rüger, R., Meindl, A., Zachau, H. G., Raßhofer, R., Roggendorf, M., Wolf, H., Arnold, N., Wienberg, J., and Kessler, C. (1991) Non-radioactive labeling and detection of nucleic acids: III. Applications of the digoxigenin system. Biol. Chem. Hoppe-Seyler 371, 939–951.CrossRefGoogle Scholar
  15. 15.
    Muhlegger, K., Huber, E., von der Eltz, H., Rüger, R., and Kessler, C. (1990) Non-radioactive labeling and detection of nucleic acids: IV. Synthesis and properties of the nucleotide compounds of the digoxigenin system and of photodig-oxigenin. Biol. Chem. Hoppe-Seyler 371, 953–965.PubMedCrossRefGoogle Scholar
  16. 16.
    Rdger, R., Höltke, H.-J., Sagner, G., Seibl, R., and Kessler, C. (1991) Rapid labelling methods using the DIG-system: incorporation of digoxigenin in PC reactions and labelling of nucleic acids with photodigoxigenin. Fresenius’ Z. Anal. Chem. 337, 114.Google Scholar
  17. 17.
    Schaap, A. P., Sandison, M. D., and Handley, R. S. (1987) Chemical and enzymatic triggering of 1,2-dioxetanes. Alkalme phosphatase-catalyzed chemilu-minescence from an aryl phosphate-substituted dioxetane. Tetrahedron Lett. 28, 1159–1162.CrossRefGoogle Scholar
  18. 18.
    Bronstein, I., Edwards, B., and Voyta, J. C. (1989) 1,2-Dioxetanes: novel chemi-luminescent enzyme substrates. Applications to immunoassay. L. Biolum. Chemolum. 4, 99–111.CrossRefGoogle Scholar
  19. 19.
    Hòltke, H. J., Sagner, G., Kessler, C., and Schmitz, G. G. (1992) Sensitive chemiluminescent detection of digoxigenin-labeled nucleic acids: A fast and simple protocol and its applications. Biotechnlques 12, 104–113.Google Scholar
  20. 20.
    Innis, M. A., Gelfand, D. H., Sninsky, J. J., and White, T. J. (1990) PCR Protocols. A Guide to Methods and Applications. Academic, New York.Google Scholar
  21. 21.
    Kessler, C. (1991) The digoxigenin: anti-digoxigenin (DIG) technology-a survey on the concept and realization of a novel bioanalytical indicator system. Mol. Cell. Probes 5, 161–205.PubMedCrossRefGoogle Scholar
  22. 22.
    Keller, G. H. and Manak, M. M. (1989) DNA Probes. Stockton, New York.Google Scholar
  23. 23.
    Kricka, L. J. (1992) Nomsotoplc DNA Probe Techmques. Academic, San Diego, CA.Google Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 1993

Authors and Affiliations

  • Udo Reischl
    • 1
  • Rüdiger Rüger
    • 1
  • Christoph Kessler
    • 1
  1. 1.Department of Molecular GeneticsBoehringer Mannheim GmbHPenzberg/Obb.Germany

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