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Related Concept Videos

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...
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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.
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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. 
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Next-generation Sequencing

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Targeted DNA Methylation Analysis by Next-generation Sequencing
08:38

Targeted DNA Methylation Analysis by Next-generation Sequencing

Published on: February 24, 2015

Determining DNA methylation profiles using sequencing.

Suhua Feng1, Liudmilla Rubbi, Steven E Jacobsen

  • 1Department of Molecular, Cell and Developmental Biology, University of California, Los Angeles, CA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|March 25, 2011
PubMed
Summary
This summary is machine-generated.

This study details two protocols for bisulfite sequencing library preparation to map DNA methylation across the entire genome. These methods enable precise determination of cytosine methylation states for comprehensive epigenetic analysis.

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DNA Methylation: Bisulphite Modification and Analysis
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DNA Methylation: Bisulphite Modification and Analysis

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

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Last Updated: Jun 3, 2026

Targeted DNA Methylation Analysis by Next-generation Sequencing
08:38

Targeted DNA Methylation Analysis by Next-generation Sequencing

Published on: February 24, 2015

DNA Methylation: Bisulphite Modification and Analysis
12:34

DNA Methylation: Bisulphite Modification and Analysis

Published on: October 21, 2011

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:

  • Epigenetics and Genomics
  • Molecular Biology

Background:

  • Cytosine methylation is a critical epigenetic mark regulating DNA transcription and replication.
  • DNA methylation patterns are heritable and crucial for cellular differentiation and tissue-specific gene expression.
  • DNA demethylases influence global methylation levels during development.

Purpose of the Study:

  • To present detailed protocols for preparing bisulfite-treated DNA libraries for next-generation sequencing.
  • To enable whole-genome analysis of cytosine methylation states.
  • To describe specialized protocols for mapping bisulfite-converted sequencing reads.

Main Methods:

  • Utilized bisulfite sequencing, the gold standard for measuring cytosine methylation.
  • Developed two distinct library preparation protocols: one with pre-methylated adapters and another with a two-stage adapter strategy.
  • Included specialized protocols for mapping bisulfite-converted sequencing reads to a reference genome.

Main Results:

  • Successfully adapted bisulfite sequencing for whole-genome methylation analysis using Illumina GAII sequencers.
  • Provided two distinct, effective protocols for bisulfite library preparation.
  • Established methods for accurate mapping of bisulfite-converted reads to determine individual cytosine methylation states.

Conclusions:

  • The described protocols facilitate comprehensive genome-wide DNA methylation profiling.
  • These methods allow for the precise determination of the methylation state of every cytosine in the genome.
  • Advances in bisulfite sequencing library preparation enable deeper insights into epigenetic regulation.