Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Next-generation Sequencing

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.

Sanger Sequencing

Sanger Sequencing

DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...

You might also read

Related Articles

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

Sort by

Same author

Human IL-34 Deficiency Primes Microglia Toward Alzheimer's Disease-Associated States.

medRxiv : the preprint server for health sciences·2026

Same author

Landscape of copy number variants in Spanish people with dementia.

NPJ genomic medicine·2026

Same author

Listening for Alzheimer's clues: machine learning analysis of multidomain speech features for cognitive impairment screening.

Frontiers in aging neuroscience·2026

Same author

Non-linear associations between sleep duration and plasma p-tau181 in the Framingham Heart Study.

Alzheimer's & dementia : the journal of the Alzheimer's Association·2026

Same author

Low overlap of plasma and CSF protein quantitative trait loci affects protein discovery for neurological disease.

Science translational medicine·2026

Same author

GWAS meta-analysis of cerebrospinal fluid Alzheimer's biomarkers reveals loci regulating lipids, brain volume and autophagy.

Nature communications·2026

Search research articles

Related Experiment Video

Updated: Jul 13, 2026

Genotyping Single Nucleotide Polymorphisms in the Mitochondrial Genome by Pyrosequencing

Genotyping Single Nucleotide Polymorphisms in the Mitochondrial Genome by Pyrosequencing

Published on: February 10, 2023

Pyrosequencing protocol using a universal biotinylated primer for mutation detection and SNP genotyping.

Jose Luis Royo¹, Manuel Hidalgo, Agustin Ruiz

¹Departamento de Genómica Estructural, Neocodex SL, Avda. Charles Darwin 6, Acc. A, 41092 Sevilla, Spain. jlroyo@neocodex.es

Nature Protocols

|July 21, 2007

Summary

This study introduces a novel method for DNA analysis using Pyrosequencing, eliminating the need for multiple biotinylated primers. This streamlined approach simplifies SNP genotyping and mutation detection, making genetic research more efficient.

More Related Videos

Pyrosequencing: A Simple Method for Accurate Genotyping

Pyrosequencing: A Simple Method for Accurate Genotyping

Published on: January 8, 2008

Pyrosequencing for Microbial Identification and Characterization

Pyrosequencing for Microbial Identification and Characterization

Published on: August 22, 2013

Related Experiment Videos

Last Updated: Jul 13, 2026

Genotyping Single Nucleotide Polymorphisms in the Mitochondrial Genome by Pyrosequencing

Genotyping Single Nucleotide Polymorphisms in the Mitochondrial Genome by Pyrosequencing

Published on: February 10, 2023

Pyrosequencing: A Simple Method for Accurate Genotyping

Pyrosequencing: A Simple Method for Accurate Genotyping

Published on: January 8, 2008

Pyrosequencing for Microbial Identification and Characterization

Pyrosequencing for Microbial Identification and Characterization

Published on: August 22, 2013

Area of Science:

Biotechnology
Genomics
Molecular Biology

Background:

DNA sequencing advances biomedical research by identifying numerous human genome polymorphisms.
Pyrosequencing is a valuable tool for medium-throughput genotyping, mutation detection, and DNA methylation analysis.
Standard Pyrosequencing requires specific biotinylated primers for each SNP, increasing complexity and cost.

Purpose of the Study:

To present an alternative protocol for Pyrosequencing that avoids the need for SNP-specific biotinylated primers.
To simplify and improve the efficiency of single nucleotide polymorphism (SNP) genotyping and other DNA analyses.

Main Methods:

Developed a protocol using a single biotinylated primer incorporated into M13-tagged PCR products.
Detailed steps for PCR amplification, single-stranded DNA template capture, and Pyrosequencing assay.
Determined optimal primer stoichiometry for the assay.

Main Results:

Successfully circumvented the requirement for SNP-specific biotinylated primers in Pyrosequencing.
The streamlined protocol allows for rapid genotyping results (15-20 min per plate) after PCR amplification.
The method is suitable for medium-throughput analysis, including SNP genotyping and mutation detection.

Conclusions:

The presented protocol offers a more efficient and cost-effective alternative to standard Pyrosequencing labeling methods.
This innovation facilitates broader application of Pyrosequencing in genetic research and diagnostics.
The simplified approach enhances the accessibility and speed of analyzing genetic variations.