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

Fluorescence-based, high-throughput DNA polymerase assay.

Liming Yu1, Guolu Hu, Leighton Howells

  • 1Amersham Biosciences, Piscataway, NJ 08855, USA. liming.yu@am.amershambiosciences.com

Biotechniques
|October 26, 2002
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

Correction: Primary diffuse large B-cell lymphoma of the lip: a case report and literature review.

Frontiers in oncology·2026
Same author

Primary diffuse large b-cell lymphoma of the lip: a case report and literature review.

Frontiers in oncology·2026
Same author

Corrigendum to "Targeting HSP90 suppresses STAT1/CCL8-driven inflammation and mitigates mitochondrial dysfunction to attenuate hypertension-induced atrial fibrillation" [Life Sci. 391 (2026) 124279].

Life sciences·2026
Same author

Craniofacial and occlusal phenotypes associated with questionnaire-defined risk of sleep-disordered breathing in children: a cross-sectional study.

BMC oral health·2026
Same author

DNA Polymerase Gamma Acetylation Governs Mitochondrial Homeostasis and Vascular Cell Senescence.

International journal of biological sciences·2026
Same author

Comparison of an Ultrasound-Based System and the Bishop Score for Predicting Labor Induction Timing in Singleton Pregnancies at Term: A Prospective Cohort Study.

International journal of women's health·2026

A new nonradioactive, fluorescence-based DNA polymerase assay offers a simpler, more stable alternative to traditional radioactive methods. This high-throughput assay provides comparable sensitivity and dynamic range for accurate enzyme activity measurement.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Assay Development

Background:

  • Traditional DNA polymerase assays rely on radioactive nucleotides, which are laborious, hazardous, and variable.
  • Existing methods present challenges for high-throughput screening and routine laboratory use.

Purpose of the Study:

  • To develop a novel, nonradioactive fluorescence-based assay for DNA polymerase activity.
  • To offer a simpler, more stable, and high-throughput alternative to radioactive assays.

Main Methods:

  • Utilized Cydye-labeled nucleotides and a biotinylated primer.
  • Employed a streptavidin-coated microplate for detection.
  • Optimized template DNA sequences to avoid consecutive incorporation of labeled nucleotides.

Related Experiment Videos

Main Results:

  • The fluorescence-based assay demonstrated sensitivity and dynamic range comparable to radioactive methods.
  • The assay proved to be simple, stable, and nonradioactive.
  • The method is suitable for high-throughput applications.

Conclusions:

  • The developed fluorescence assay is a viable, advantageous alternative to radioactive DNA polymerase assays.
  • This method enhances efficiency, safety, and throughput in enzyme activity measurement.
  • Careful selection of template DNA sequences is crucial for optimal assay performance.