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Next-generation Sequencing

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....

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Updated: Mar 15, 2026

Targeted DNA Methylation Analysis by Next-generation Sequencing

Targeted DNA Methylation Analysis by Next-generation Sequencing

Published on: February 24, 2015

DNA methylation detection at single base resolution using targeted next generation bisulfite sequencing and cross

Smitha Bhat¹, Sandeep Mallya¹, Vinay Koshy Varghese¹

¹Department of Biotechnology, School of Life Sciences, Manipal University, Manipal 576104, Karnataka, India.

|September 18, 2016

Summary

This study introduces a rapid method for DNA methylation detection using targeted next-generation bisulfite sequencing. The technique accurately quantifies methylation across multiple gene loci and samples, aiding biomarker discovery.

Keywords:

Biomarker CpG islands DNA methylation Gene discovery Next generation sequencing (NGS)

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Last Updated: Mar 15, 2026

Targeted DNA Methylation Analysis by Next-generation Sequencing

Targeted DNA Methylation Analysis by Next-generation Sequencing

Published on: February 24, 2015

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

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

Published on: February 24, 2015

Methyl-binding DNA capture Sequencing for Patient Tissues

Methyl-binding DNA capture Sequencing for Patient Tissues

Published on: October 31, 2016

Area of Science:

Epigenetics
Genomics
Molecular Biology

Background:

DNA methylation is a crucial epigenetic modification regulating gene expression.
Accurate and simultaneous determination of DNA methylation across multiple sites and samples is challenging.
Understanding DNA methylation patterns is vital for biomarker identification and disease research.

Purpose of the Study:

To develop and demonstrate a rapid, accurate method for simultaneous DNA methylation detection.
To analyze DNA methylation in multiple CpG sites across 12 gene loci in human peripheral blood DNA.
To validate the method's reliability and efficiency for large-scale epigenetic studies.

Main Methods:

Targeted next-generation bisulfite sequencing was employed.
Genomic DNA from 14 healthy individuals was analyzed.
DNA methylation was quantified at 465 CpG sites across 12 gene loci.

Main Results:

The method successfully detected DNA methylation in multiple loci and samples simultaneously.
Significant methylation levels were observed in BCR (25.6%), IL17RA (89.2%), and RBM38 (38.9%).
Technical validation using direct capillary sequencing confirmed high correlation and agreement with the new method.

Conclusions:

Targeted next-generation bisulfite sequencing offers a fast and reliable approach for quantitative DNA methylation analysis.
The method provides single-base resolution and good coverage, suitable for biomarker discovery.
This technique facilitates exploration of gene panels for biomarker identification and functional validation.