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

Consecutive DNA terminator sequencing by using enzymatically generated primers.

M Speek, H Ilves, A Lind

    Analytical Biochemistry
    |November 1, 1986
    PubMed
    Summary
    This summary is machine-generated.

    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

    Critical COVID-19 is associated with distinct leukocyte phenotypes and transcriptome patterns.

    Journal of internal medicine·2021
    Same author

    Increased inflammatory markers reflecting fibrogenesis are independently associated with cardiac involvement in hospitalized COVID-19 patients.

    The Journal of infection·2021
    Same author

    Elevated markers of gut leakage and inflammasome activation in COVID-19 patients with cardiac involvement.

    Journal of internal medicine·2020
    Same author

    Distinct and early increase in circulating MMP-9 in COVID-19 patients with respiratory failure.

    The Journal of infection·2020
    Same author

    Acupressure treatment for methotrexate-induced nausea in arthritis patients.

    Scandinavian journal of rheumatology·2019
    Same author

    [Infective endocarditis : New ESC guidelines 2015].

    Der Internist·2016
    Same journal

    Lysozyme assay using a rationally designed GN4G2 substrate with coupled β-glucosidase reaction.

    Analytical biochemistry·2026
    Same journal

    The long run: A tribute to Arthur Joseph Lawrence Cooper.

    Analytical biochemistry·2026
    Same journal

    Evaluation of a method for affinity measurement using solution equilibrium titration with magnetic beads.

    Analytical biochemistry·2026
    Same journal

    Metabolomics approach using UHPLC/QE-MS for the mechanism of He Xue Ming Mu tablets on non-proliferative diabetic retinopathy.

    Analytical biochemistry·2026
    Same journal

    UniRES-GO: Unified residue-level early fusion of sequence and predicted structure for protein function prediction.

    Analytical biochemistry·2026
    Same journal

    IgG detection by enzyme-linked mass spectrometric assay versus color, fluorescent, ECL in buffer and serum.

    Analytical biochemistry·2026
    See all related articles

    This study presents an improved DNA sequencing strategy using enzymatically generated primers for M13 recombinant DNA. This method enables efficient determination of long DNA sequences up to 6 kb.

    Area of Science:

    • Molecular Biology
    • Genomics
    • Biotechnology

    Background:

    • Dideoxy chain termination sequencing is a fundamental technique in molecular biology.
    • Generating specific primers is crucial for efficient and accurate DNA sequencing.
    • Existing methods for primer generation can be inefficient or lack specificity.

    Purpose of the Study:

    • To develop an improved strategy for dideoxy chain termination sequencing of M13 recombinant DNA.
    • To utilize enzymatically generated primers for enhanced sequencing efficiency and accuracy.
    • To enable the determination of longer DNA sequences through a streamlined process.

    Main Methods:

    • Synchronous extension of a universal primer using the Klenow fragment of DNA polymerase I.
    • Enzymatic cleavage of extended primers at preselected restriction enzyme sites to generate homogeneous 5' ends.

    Related Experiment Videos

  • Iterative application of primer extension and cleavage, guided by microcomputer analysis for restriction site selection.
  • Sequencing of M13 recombinant DNA using the generated primers.

    • Main Results:

    • Successfully generated short primers with homogeneous 5' ends for dideoxy sequencing.
    • Determined DNA sequences approximately 6 kb long from multiple M13 recombinant DNA samples.
    • Utilized twenty different restriction enzymes to facilitate the primer generation and sequencing process.
    • Demonstrated the efficiency and robustness of the primer extension-cleavage-sequencing strategy.

    Conclusions:

    • The presented strategy offers an improved method for dideoxy chain termination sequencing of M13 recombinant DNA.
    • Enzymatically generated primers provide a reliable and efficient means for extending DNA sequence determination.
    • This approach facilitates the sequencing of large DNA fragments, advancing genomic research capabilities.