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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.
Sanger Sequencing01:57

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...
RNA-seq03:21

RNA-seq

RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases. 
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...
Single Nucleotide Polymorphisms-SNPs01:05

Single Nucleotide Polymorphisms-SNPs

A single nucleotide polymorphism or SNP is a single nucleotide variation at a specific genomic position in a large population. It is the most prevalent type of sequence variation found in the human genome. Point mutations that occur in more than 1% of the population qualify as SNPs. These are present once every 1000 nucleotides on an average in the human genome. Replacement of a purine with another purine (A/G) or a pyrimidine with another pyrimidine (C/T) is known as a transition. In contrast,...

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Updated: Jun 20, 2026

Pyrosequencing: A Simple Method for Accurate Genotyping
13:06

Pyrosequencing: A Simple Method for Accurate Genotyping

Published on: January 8, 2008

Pyrosequencing for SNP genotyping.

Jose Luis Royo1, Jose Jorge Galán

  • 1Department of Structural Genomics, Neocodex SL, Seville, Spain.

Methods in Molecular Biology (Clifton, N.J.)
|September 22, 2009
PubMed
Summary
This summary is machine-generated.

Pyrosequencing is a real-time DNA sequencing technology that measures light produced by nucleotide incorporation. This method enables quantitative analysis for applications like single-nucleotide polymorphism genotyping.

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Published on: August 22, 2013

Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genetics

Background:

  • Pyrosequencing is a real-time DNA sequencing method.
  • It relies on detecting light signals generated during DNA synthesis.

Purpose of the Study:

  • To describe the pyrosequencing process.
  • To highlight its application in single-nucleotide polymorphism (SNP) genotyping.
  • To showcase other potential applications.

Main Methods:

  • DNA polymerase incorporates nucleotides, releasing pyrophosphates.
  • Sulfurylase converts pyrophosphates to ATP.
  • Luciferase uses ATP to produce light, detected by a CCD camera.

Main Results:

  • Quantitative measurement of incorporated nucleotides is possible.
  • The reaction is rapid (milliseconds).
  • The method is adaptable for various genetic analyses.

Conclusions:

  • Pyrosequencing offers a versatile platform for DNA analysis.
  • It is particularly effective for SNP genotyping.
  • Applications extend to loss of heterozygosity and methylation studies.