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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.
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...
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...
Maxam-Gilbert Sequencing01:05

Maxam-Gilbert Sequencing

In the same year as the discovery of the Sanger sequencing method, another group of scientists, Allan Maxam and Walter Gilbert, demonstrated their chemical-cleavage method for DNA sequencing. The Maxam-Gilbert method relies on using different chemicals that can cleave the DNA sequence at specific sites, the separation of resulting DNA fragments of variable size using electrophoresis, and deciphering the DNA sequence from the resulting gel bands.
Challenges of the Maxam-Gilbert Method
The...
PCR01:32

PCR

Overview
PCR - Polymerase Chain Reaction01:32

PCR - Polymerase Chain Reaction

Overview

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Related Experiment Video

Updated: May 8, 2026

Pyrosequencing: A Simple Method for Accurate Genotyping
13:06

Pyrosequencing: A Simple Method for Accurate Genotyping

Published on: January 8, 2008

Fundamentals of pyrosequencing.

Colleen T Harrington1, Elaine I Lin, Matthew T Olson

  • 1Sol Goldman Pancreatic Cancer Research Center, Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21231, USA.

Archives of Pathology & Laboratory Medicine
|September 3, 2013
PubMed
Summary
This summary is machine-generated.

Pyrosequencing offers a simpler, more sensitive DNA sequencing method than Sanger sequencing for detecting genetic mutations. This technique analyzes pyrophosphate release to determine DNA sequences, aiding in genetic disorder identification.

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Genotyping Single Nucleotide Polymorphisms in the Mitochondrial Genome by Pyrosequencing

Published on: February 10, 2023

Area of Science:

  • Molecular Biology
  • Genetics
  • Bioinformatics

Background:

  • DNA sequencing is crucial for identifying genetic disorders and cancers caused by DNA mutations.
  • Pyrosequencing presents advantages over Sanger sequencing, including reduced complexity, fewer steps, and a superior limit of detection.
  • The core principle of pyrosequencing involves monitoring pyrophosphate release during deoxyribonucleotide triphosphate incorporation into DNA.

Purpose of the Study:

  • To elucidate the fundamental principles and applications of pyrosequencing technology.
  • To demonstrate the practical use of pyrosequencing for analyzing DNA sequences and mutations.

Main Methods:

  • Utilized the free software Pyromaker to simulate and visualize pyrosequencing results.
  • Input DNA sequences and pyrosequencing parameters into Pyromaker to generate expected pyrograms.
  • Analyzed pyrograms to differentiate between wild-type and mutant DNA sequences.

Main Results:

  • Demonstrated distinct pyrogram patterns for mutant versus wild-type DNA sequences.
  • Illustrated the analysis of pyrogram peaks corresponding to specific mutations in tumor DNA.
  • Identified limitations, such as the potential indistinguishability of complex mutations from single base changes.

Conclusions:

  • Pyrosequencing provides a valuable method for DNA sequence analysis and mutation detection.
  • The principles of pyrosequencing underpin advanced next-generation sequencing platforms like Roche 454 and Ion Torrent.
  • Understanding pyrogram analysis is key to interpreting sequencing data and identifying genetic variations.