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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...
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...

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Related Experiment Video

Updated: May 9, 2026

Pyrosequencing for Microbial Identification and Characterization
12:37

Pyrosequencing for Microbial Identification and Characterization

Published on: August 22, 2013

Bisulfite pyrosequencing.

Christopher F Bassil1, Zhiqing Huang, Susan K Murphy

  • 1Division of Gynecologic Oncology, Duke University Medical Center, Durham, NC, USA.

Methods in Molecular Biology (Clifton, N.J.)
|August 6, 2013
PubMed
Summary
This summary is machine-generated.

Bisulfite pyrosequencing quantifies DNA methylation at individual CG sites within short DNA regions. This sequencing-by-synthesis method offers a detailed methylation profile for PCR amplicons.

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Enhanced Reduced Representation Bisulfite Sequencing for Assessment of DNA Methylation at Base Pair Resolution

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Last Updated: May 9, 2026

Pyrosequencing for Microbial Identification and Characterization
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Pyrosequencing for Microbial Identification and Characterization

Published on: August 22, 2013

Pyrosequencing: A Simple Method for Accurate Genotyping
13:06

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Enhanced Reduced Representation Bisulfite Sequencing for Assessment of DNA Methylation at Base Pair Resolution
13:47

Enhanced Reduced Representation Bisulfite Sequencing for Assessment of DNA Methylation at Base Pair Resolution

Published on: February 24, 2015

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • DNA methylation is a crucial epigenetic modification regulating gene expression.
  • Accurate quantification of DNA methylation is essential for understanding various biological processes and diseases.
  • Existing methods may have limitations in resolution or throughput for specific applications.

Purpose of the Study:

  • To describe the bisulfite pyrosequencing method for quantitative DNA methylation analysis.
  • To highlight its application in determining methylation status of individual CG cytosines.
  • To present its utility for analyzing PCR amplicons up to 115 bases.

Main Methods:

  • Bisulfite conversion of DNA to differentiate methylated (cytosine) from unmethylated (thymine) CG sites.
  • Sequencing-by-synthesis approach involving stepwise nucleotide incorporation.
  • Detection of pyrophosphate release, proportional to nucleotide incorporation, via light emission.

Main Results:

  • Quantitative determination of methylation at individual CG cytosines within PCR amplicons.
  • Generation of a methylation profile for the targeted DNA region.
  • The method provides a quantitative portrait of methylation status.

Conclusions:

  • Bisulfite pyrosequencing is an effective method for precise DNA methylation quantification.
  • It enables detailed analysis of methylation patterns in specific DNA regions.
  • This technique is valuable for epigenetic research and diagnostics.