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Hybridization and Detection of Fluorescein-Labeled DNA Probes Using Chemiluminescence

  • Claire B. Wheeler
  • Bronwen M. Harvey
  • Martin Cunningham
Protocol
  • 126 Downloads
Part of the Springer Protocols Handbooks book series (SPH)

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.

Keywords

Sodium Dodecyl Sulfate Amersham Pharmacia Biotech Dextran Sulfate Standard Saline Citrate Stringency Wash 
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References

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

Copyright information

© Humana Press Inc., Totowa, NJ 2000

Authors and Affiliations

  • Claire B. Wheeler
    • 1
  • Bronwen M. Harvey
    • 1
  • Martin Cunningham
    • 1
  1. 1.Research and DevelopmentAmersham Pharmacia Biotech, Ltd.UK

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