Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Using DNA fragments as probes.

W M Strauss1

  • 1Harvard Medical School, Boston, Massachusetts, USA.

Current Protocols in Molecular Biology
|February 12, 2008
PubMed
Summary
This summary is machine-generated.

This study details a method for detecting specific DNA sequences in bacteriophage plaques or bacterial colonies using radioactive probes and filter hybridization. The protocol optimizes DNA reannealing near the melting temperature (Tm) for accurate clone identification.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Preparation of genomic DNA from mammalian tissue.

Current protocols in immunology·2008
Same author

Purification and characterization of YACs containing large inserts.

Current protocols in human genetics·2008
Same author

Preparation of genomic DNA from mammalian tissue.

Current protocols in neuroscience·2008
Same author

Preparation of genomic DNA from mammalian tissue.

Current protocols in molecular biology·2008
Same author

Non-disruptive PNA-FISH protocol for formalin-fixed and paraffin-embedded tissue sections.

BioTechniques·2001
Same author

PNA interference mapping demonstrates functional domains in the noncoding RNA Xist.

Proceedings of the National Academy of Sciences of the United States of America·2001
Same journal

Nondenaturing Polyacrylamide Gel Electrophoresis: Preparation and Analysis of DNA.

Current protocols in molecular biology·2021
Same journal

Purification and Concentration of DNA from Aqueous Solutions: Preparation and Analysis of DNA.

Current protocols in molecular biology·2021
Same journal

Expression of Proteins Using Semliki Forest Virus Vectors: Protein Expression.

Current protocols in molecular biology·2021
Same journal

Methylation and Uracil Interference Assays for Analysis of Protein-DNA Interactions: DNA-Protein Interactions.

Current protocols in molecular biology·2021
Same journal

Separation of Double- and Single-Stranded Nucleic Acids Using Hydroxylapatite Chromatography: Preparation and Analysis of DNA.

Current protocols in molecular biology·2021
Same journal

Pulsed-Field Gel Electrophoresis: Preparation and Analysis of DNA.

Current protocols in molecular biology·2021
See all related articles

Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Nucleic acid hybridization is fundamental to molecular biology techniques.
  • Efficient detection of specific DNA sequences in biological samples is crucial for research and diagnostics.
  • Optimizing hybridization conditions, such as temperature near the melting temperature (Tm), enhances specificity and sensitivity.

Purpose of the Study:

  • To describe a detailed procedure for detecting bacteriophage plaques and bacterial colonies using DNA hybridization.
  • To enable the identification of specific clones through hybridization with radioactive probes.
  • To present an alternative protocol that omits formamide from the hybridization solution.

Main Methods:

  • Bacteriophage plaques or bacterial colonies are immobilized on a filter membrane.

Related Experiment Videos

  • Filters are prewet and placed in a sealed bag for hybridization with a radioactive DNA probe.
  • Post-hybridization washes remove excess probe, followed by autoradiography for clone visualization.

    • Main Results:

    • The described hybridization method successfully identifies clones containing specific DNA sequences.
    • Autoradiography allows for the precise localization of positive signals on the filter membrane.
    • An alternate protocol provides a variation in hybridization solution composition.

    Conclusions:

    • This hybridization technique provides a reliable method for screening and identifying specific DNA sequences in microbial colonies.
    • The protocol is adaptable, with an alternative method offered for different experimental needs.
    • The method facilitates the discovery and characterization of desired clones in molecular biology research.