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

DNA sequencing using differential extension with nucleotide subsets (DENS)

M C Raja1, D Zevin-Sonkin, J Shwartzburd

  • 1Center for Mechanistic Biology and Biotechnology, Argonne National Laboratory IL 60439-4833, USA.

Nucleic Acids Research
|February 15, 1997
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

Predicting band gap from chemical composition: a comparative study of machine learning and deep learning models.

Scientific reports·2026
Same author

Cluster-size-dependent cooperativity in sulfur-centred hydrogen bonds: ethane-1,2-dithiol with HF, H<sub>2</sub>O, and NH<sub>3</sub>.

Physical chemistry chemical physics : PCCP·2026
Same author

Total antibodies and neutralizing ability of convalescent sera against three different strains of SARS-CoV-2.

Voprosy virusologii·2025
Same author

First detection of influenza A virus subtypes H1N1 and H3N8 in the Antarctic region: King George Island, 2023.

Voprosy virusologii·2024
Same author

Combined FTIR/Raman spectroscopic studies and ab initio electronic structure calculations of Dithiothreitol.

Spectrochimica acta. Part A, Molecular and biomolecular spectroscopy·2023
Same author

Aerosol demasking enhances climate warming over South Asia.

NPJ climate and atmospheric science·2023

Differential Extension with Nucleotide Subsets (DENS) enables enzymatic primer synthesis, bypassing chemical steps for robust DNA sequencing. This novel method enhances primer extension selectivity, improving primer walking efficiency and reliability.

Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genomics

Background:

  • Primer walking is crucial for DNA sequencing but often relies on chemically synthesized primers.
  • Existing primer synthesis methods can be inefficient and costly, limiting scalability.

Purpose of the Study:

  • To introduce a novel enzymatic primer synthesis technique, Differential Extension with Nucleotide Subsets (DENS).
  • To demonstrate DENS's ability to avoid chemical synthesis steps in primer walking.
  • To enhance the robustness and efficiency of DNA sequencing.

Main Methods:

  • Selective enzymatic extension of short primers using limited deoxynucleotide triphosphate (dNTP) subsets at specific temperatures.
  • Utilizing a subsequent termination reaction to differentiate correctly extended primers from off-target extensions.

Related Experiment Videos

  • Employing longer, degenerate primers (up to 8-mers) compatible with thermostable polymerases.

    • Main Results:

    • DENS achieves selective primer extension of 6-9 bases or longer at intended sites.
    • The method effectively distinguishes intended primers from shorter, off-target extensions.
    • DENS enables the use of stronger-priming, longer primers with thermostable polymerases.

    Conclusions:

    • DENS offers a template-directed enzymatic approach to primer synthesis, eliminating chemical steps.
    • This technique improves primer walking efficiency and robustness, particularly for double-stranded DNA sequencing.
    • DENS is compatible with cycle sequencing using dye-terminators and Taq DNA polymerase.