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 Concept Videos

Next-generation Sequencing03:00

Next-generation Sequencing

The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features.

You might also read

Related Articles

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

Sort by
Same author

Determinants of DNA-sequence-based Diagnostic Yield in the CSER Consortium.

medRxiv : the preprint server for health sciences·2026
Same author

Simultaneous detection of small and large variants enhances the diagnosis of rare diseases using full genome sequencing.

Human molecular genetics·2026
Same author

The Taiwan Precision Medicine Initiative provides a cohort for large-scale studies.

Nature·2025
Same author

Population-specific polygenic risk scores for people of Han Chinese ancestry.

Nature·2025
Same author

Clinical impact of pharmacogenetic risk variants in a large chinese cohort.

Nature communications·2025
Same author

Identification of De Novo Chromosomal Translocations Disrupting NIPBL in a Patient With Cornelia de Lange Syndrome by Full Genome Analysis.

Molecular genetics & genomic medicine·2025

Related Experiment Video

Updated: Jul 5, 2026

Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions
08:23

Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions

Published on: September 25, 2018

High-throughput genotyping with primer extension fluorescent polarization detection.

Pui-Yan Kwok1

  • 1University of California, San Francisco, California, USA.

Current Protocols in Human Genetics
|April 23, 2008
PubMed
Summary

Primer extension assays with fluorescence polarization (FP) detection offer a versatile method for single nucleotide polymorphism (SNP) genotyping. This homogeneous assay simplifies genetic analysis by detecting nucleotide incorporation without sample purification.

More Related Videos

Candidate Gene Testing in Clinical Cohort Studies with Multiplexed Genotyping and Mass Spectrometry
05:53

Candidate Gene Testing in Clinical Cohort Studies with Multiplexed Genotyping and Mass Spectrometry

Published on: June 21, 2018

Multiplexed Isothermal Amplification Based Diagnostic Platform to Detect Zika, Chikungunya, and Dengue 1
06:18

Multiplexed Isothermal Amplification Based Diagnostic Platform to Detect Zika, Chikungunya, and Dengue 1

Published on: March 13, 2018

Related Experiment Videos

Last Updated: Jul 5, 2026

Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions
08:23

Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions

Published on: September 25, 2018

Candidate Gene Testing in Clinical Cohort Studies with Multiplexed Genotyping and Mass Spectrometry
05:53

Candidate Gene Testing in Clinical Cohort Studies with Multiplexed Genotyping and Mass Spectrometry

Published on: June 21, 2018

Multiplexed Isothermal Amplification Based Diagnostic Platform to Detect Zika, Chikungunya, and Dengue 1
06:18

Multiplexed Isothermal Amplification Based Diagnostic Platform to Detect Zika, Chikungunya, and Dengue 1

Published on: March 13, 2018

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Single nucleotide polymorphism (SNP) genotyping is crucial for genetic studies.
  • Traditional genotyping methods often require complex procedures like separation and purification.
  • A homogeneous assay that simplifies detection is highly desirable.

Purpose of the Study:

  • To present the primer extension assay with fluorescence polarization (FP) detection as a versatile tool for SNP genotyping.
  • To highlight the advantages of this homogeneous assay under universal conditions.
  • To explain the principle of FP detection in the context of primer extension.

Main Methods:

  • Utilizing a primer extension assay where DNA polymerase incorporates an allelic nucleotide.
  • Employing fluorescence polarization (FP) detection to monitor the reaction.
  • Designing primers to anneal one base upstream of the polymorphic site on target DNA.

Main Results:

  • Fluorescence polarization (FP) occurs when a fluorescent dye attached to a large molecule is excited by polarized light.
  • Incorporation of a fluorescently labeled nucleotide results in a significant increase in FP.
  • This increase in FP allows for direct determination of the incorporated nucleotide and thus the genotype.

Conclusions:

  • The primer extension assay with FP detection is a robust and homogeneous method for SNP genotyping.
  • This assay simplifies the process by eliminating the need for post-reaction separation or purification.
  • The method provides a straightforward way to determine DNA sample genotypes based on nucleotide incorporation.