Skip to main content

Biotinylated Probes in Colony Hybridization

  • Protocol
  • 920 Accesses

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 80))

Abstract

Colony hybridization is a procedure that allows the detection of cells containing nucleic acid sequences of interest (1). In this method, microbial colonies grown on, or transferred to, a supporting membrane are lysed and their nucleic acids denatured to single strands and fixed in place on the membrane. The membrane is then exposed to a similarly denatured “probe” sequence, which is identical or homologous to all or part of the target sequence, under conditions favoring reanneahng. Probe sequences hybridize to complementary sequences on the membrane. Positive hybridization events are then detected by determining the presence and location of probe sequences on the membrane

This is a preview of subscription content, log in via an institution.

Buying options

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   74.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Grunstein, M and Hogness, D. S (1975) Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc. Natl. Acad Sci USA 72, 3961–3965.

    Article  PubMed  CAS  Google Scholar 

  2. Langer, P. R., Waldrop, A. A., and Ward, D. C. (1981) Enzymatic synthesis of biotin-labeled polynucleotides: novel nucleic acid affinity probes. Proc Natl. Acad. Sci USA 78, 6633–6637

    Article  PubMed  CAS  Google Scholar 

  3. Brigati, D. J., Myerson, D., Leary, J. J., Spalholz, B., Travis, S Z., Fong, D. K. Y., Hsiung, G. D., and Ward, D. C. (1983) Detection of viral genomes in cultured cells and paraffin-embedded tissue sections using biotin-labeled hybridization. Virology 126, 32–50

    Article  PubMed  CAS  Google Scholar 

  4. Boffey, S. A. (1984) Plasmid DNA isolation by the cleared lysate method, in Methods in Molecular Biology, vol 2: Nucleic Acids (Walker, J M., ed.), Humana, Clifton, NJ, pp. 177–183.

    Google Scholar 

  5. Maas, R. (1983) An improved colony hybridization method with significantly increased sensitivity for detection of single genes Plasmid 10, 296–298.

    Article  PubMed  CAS  Google Scholar 

  6. Forster, A C, McInnes, J L., Skingle, D. C, and Symons, R H (1985) Nonradioactive hybridization probes prepared by the chemical labeling of DNA and RNA with a novel reagent, photobiotin Nucleic Acids Res 13, 745–761.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 1998 Humana Press Inc., Totowa, NJ

About this protocol

Cite this protocol

Haas, M.J. (1998). Biotinylated Probes in Colony Hybridization. In: Pound, J.D. (eds) Immunochemical Protocols. Methods in Molecular Biology™, vol 80. Humana Press. https://doi.org/10.1007/978-1-59259-257-9_40

Download citation

  • DOI: https://doi.org/10.1007/978-1-59259-257-9_40

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-493-8

  • Online ISBN: 978-1-59259-257-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics