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The Nucleic Acid Protocols Handbook pp 153–156Cite as

Hybridization and Detection of Fluorescein-Labeled DNA Probes Using Chemiluminescence

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Abstract

DNA probes can be labeled using a fluorescein-labeled nucleotide and subsequently detected with a dioxetane-based chemiluminescence system using an antifluorescein antibody conjugated to alkaline phosphatase (1,2). The use of chemiluminescence avoids the fading of results associated with some colorimetric detection procedures used with some nonradioactive labeling and detection systems. Hard-copy results showing excellent resolution are produced.

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References

  1. Bronstein, I., Edwards, B., and Voyta, J. C. (1989) 1,2-Dioxetanes: novel chemiluminescent enzyme substrates; applications to immunoassays. J. Biolum. Chemilum. 4, 99–111.

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  2. Bronstein, I., Olesen, C. E. M., Martin, C. S., Schneider, G., Edwards, B., Sparks, A., and Voyta, J. C. (1994) Chemiluminescent detection of DNA and protein with CDP™ and CDP-STAR™ 1,2-dioxetane substrates, in Bioluminescence and Chemiluminescence: Fundamentals and Applied Aspects (Campbell, A. K., Kricka, L. J., and Stanley, P. E., eds.), Wiley, Chichester, UK, pp. 269–272.

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  3. Feinberg, A. P. and Vogelstein, B. (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal. Biochem. 132, 6–13.

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Author information

Authors and Affiliations

  1. Research and Development, Amersham Pharmacia Biotech, Ltd., Buckinghamshire, UK

    Claire B. Wheeler, Bronwen M. Harvey & Martin Cunningham

Authors
  1. Claire B. Wheeler
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  2. Bronwen M. Harvey
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  3. Martin Cunningham
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Editor information

Editors and Affiliations

  1. University of Hertfordshire, Hatfield, UK

    Ralph Rapley

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© 2000 Humana Press Inc., Totowa, NJ

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Wheeler, C.B., Harvey, B.M., Cunningham, M. (2000). Hybridization and Detection of Fluorescein-Labeled DNA Probes Using Chemiluminescence. In: Rapley, R. (eds) The Nucleic Acid Protocols Handbook. Springer Protocols Handbooks. Humana Press, Totowa, NJ. https://doi.org/10.1385/1-59259-038-1:153

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  • DOI: https://doi.org/10.1385/1-59259-038-1:153

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-0-89603-459-4

  • Online ISBN: 978-1-59259-038-4

  • eBook Packages: Springer Book Archive

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