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Related Concept Videos

Next-generation Sequencing03:00

Next-generation Sequencing

100.7K
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....
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Sanger Sequencing01:57

Sanger Sequencing

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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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RNA-seq03:21

RNA-seq

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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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Related Experiment Video

Updated: Mar 18, 2026

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

Targeted DNA Methylation Analysis by Next-generation Sequencing

Published on: February 24, 2015

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NGS-based deep bisulfite sequencing.

Suman Lee1, Joomyeong Kim1

  • 1Department of Biological Sciences, Louisiana State University, Baton Rouge, LA 70803, USA.

Methodsx
|July 2, 2016
PubMed
Summary
This summary is machine-generated.

We developed a new NGS deep bisulfite sequencing protocol for genome-wide DNA methylation analysis. This method efficiently measures DNA methylation levels and patterns at individual loci.

Keywords:
Bisulfite sequencingDNA methylationEpigenomesNGSNGS, Next-Generation-SequencingNGS-based deep bisulfite sequencingPCR, polymerase chain reaction

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Optimized Analysis of DNA Methylation and Gene Expression from Small, Anatomically-defined Areas of the Brain
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Area of Science:

  • Genomics
  • Epigenetics
  • Molecular Biology

Background:

  • DNA methylation is a critical epigenetic modification influencing gene expression.
  • Accurate and comprehensive analysis of DNA methylation patterns is essential for understanding various biological processes and diseases.
  • Existing methods for DNA methylation analysis have limitations in efficiency and resolution.

Purpose of the Study:

  • To develop a novel Next-Generation Sequencing (NGS)-based deep bisulfite sequencing protocol.
  • To enable rapid and efficient DNA methylation analysis at the genomic level.
  • To provide high-resolution data on DNA methylation levels and patterns for individual loci.

Main Methods:

  • Development of an NGS-ready library construction protocol involving individual amplification of PCR products from bisulfite-converted DNA.
  • Implementation of a bioinformatics strategy for sorting raw sequence reads generated by NGS platforms.
  • Derivation of DNA methylation levels and patterns for individual genomic loci.

Main Results:

  • The protocol enables efficient generation of NGS-ready libraries from a large number of amplified bisulfite-converted DNA fragments.
  • The bioinformatics strategy effectively sorts NGS sequence reads for accurate methylation analysis.
  • The deep bisulfite sequencing approach successfully provides quantitative DNA methylation levels and reveals informative DNA methylation patterns at individual loci, surpassing existing methods.

Conclusions:

  • The developed NGS-based deep bisulfite sequencing protocol offers an efficient and powerful tool for comprehensive DNA methylation analysis.
  • This method provides unprecedented insights into DNA methylation patterns at a high resolution.
  • The protocol is valuable for advancing research in epigenetics, genomics, and disease mechanisms.