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

Rapid and efficient cosmid cloning.

D Ish-Horowicz, J F Burke

    Nucleic Acids Research
    |July 10, 1981
    PubMed
    Summary
    This summary is machine-generated.

    This study introduces a cosmid cloning method for efficient DNA fragment insertion (32-45kb). The technique prevents self-ligation and multiple inserts, yielding high numbers of single-fragment clones from Drosophila DNA.

    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

    Delta4, an endothelial specific notch ligand expressed at sites of physiological and tumor angiogenesis.

    Differentiation; research in biological diversity·2002
    Same author

    Conserved signals and machinery for RNA transport in Drosophila oogenesis and embryogenesis.

    Nature·2001
    Same author

    View from the past.

    Surgery·2001
    Same author

    The initial attitudes of patients toward longer maintenance hemodialysis.

    Clinical nephrology·2001
    Same author

    Neuronal expression of an FMRFamide-gated Na+ channel and its modulation by acid pH.

    The Journal of neuroscience : the official journal of the Society for Neuroscience·2001
    Same author

    Mae mediates MAP kinase phosphorylation of Ets transcription factors in Drosophila.

    Nature·2001
    Same journal

    Correction to 'scSuperAnnotator: A platform for benchmarking comparison and visualizing automated cellular annotation methods for scRNA-seq data'.

    Nucleic acids research·2026
    Same journal

    Correction to 'Differentiable partition function calculation for RNA'.

    Nucleic acids research·2026
    Same journal

    Deployment of non-canonical splicing in tunicate genomes is mediated by divergent U2AF function and changing m6A modification in U1 and U6 snRNA.

    Nucleic acids research·2026
    Same journal

    Bacillus subtilis DnaB forms multiple protein-protein interactions essential for DNA replication initiation.

    Nucleic acids research·2026
    Same journal

    Multiple forms of protein-protein and DNA binding are exhibited by BrxC from the BREX phage restriction system.

    Nucleic acids research·2026
    Same journal

    Biosynthesis of glycosylated 5-hydroxycytosine in the DNA of diverse viruses.

    Nucleic acids research·2026
    See all related articles

    Area of Science:

    • Molecular Biology
    • Genetics
    • Biotechnology

    Background:

    • Cosmid cloning is essential for genomic research, enabling the study of large DNA fragments.
    • Existing methods often face challenges with self-ligation, non-contiguous fragment insertion, and insert size selection.
    • Efficient preparation of plasmid and cosmid DNA is crucial for downstream applications.

    Purpose of the Study:

    • To develop a rapid and efficient cosmid cloning procedure for individual DNA fragments.
    • To create a cloning vector system that prevents self-ligation and the formation of undesirable clones.
    • To optimize DNA fragment preparation and cloning for high-yield genomic analysis.

    Main Methods:

    • A modified cloning vector, pJb8, was engineered with specific end modifications to prevent self-ligation.

    Related Experiment Videos

  • Insert DNA fragments were generated by partial digestion and dephosphorylated to ensure single-fragment insertion.
  • The protocol was tested using Drosophila DNA, with subsequent plasmid and cosmid DNA preparation methods described.

    • Main Results:

    • The developed cosmid cloning method efficiently clones individual DNA fragments ranging from 32kb to 45kb.
    • The procedure prevents the ligation of non-contiguous fragments and eliminates the need for insert size selection.
    • Approximately 5 x 10^5 clones were obtained per microgram of Drosophila DNA, with an average insert size of 38kb.

    Conclusions:

    • This novel cosmid cloning technique offers a significant improvement in efficiency and specificity for large DNA fragment cloning.
    • The method simplifies the cloning process by preventing common artifacts and reducing the need for extensive screening.
    • The described protocol is valuable for genomic library construction and large-scale DNA manipulation in molecular biology research.