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Updated: May 12, 2026
08:38
Targeted DNA Methylation Analysis by Next-generation Sequencing
Published on: February 24, 2015
Targeted next-generation sequencing techniques: From primer design to sophisticated library construction
Maxim Koryukov1, Regina Mikheeva1, Andrey Kechin1
1Institute of Chemical Biology and Fundamental Medicine, Novosibirsk 630090, Russia; Novosibirsk State University, Novosibirsk 630090, Russia.
Methods (San Diego, Calif.)
|May 10, 2026
Summary
Targeted Next-Generation Sequencing (tNGS) relies on target enrichment, a complex step reviewed here. This analysis covers PCR and hybridization methods, computational challenges, and novel techniques for molecular diagnostics.
Area of Science:
- Molecular Biology
- Genomics
- Clinical Diagnostics
Background:
- Targeted Next-Generation Sequencing (tNGS) is crucial for clinical diagnostics and research.
- tNGS offers a cost-effective alternative to whole-genome sequencing.
- Target enrichment is the most complex and variable stage in the tNGS workflow.
Purpose of the Study:
- To critically examine PCR-based and hybridization-based target enrichment strategies for tNGS.
- To survey established and emerging enrichment techniques.
- To highlight the suitability of enrichment methods for molecular research and diagnostics.
Main Methods:
- Review of PCR-based enrichment strategies.
- Review of hybridization-based enrichment strategies.
- Analysis of computational challenges in tNGS assay design.
Main Results:
- Both PCR and hybridization methods have distinct advantages and limitations.
- Sophisticated approaches like UMIs and non-canonical bases are emerging.
- Suitability varies across different research and diagnostic applications.
Conclusions:
- Understanding enrichment methodologies is key to advancing tNGS applications.
- Addressing computational challenges is vital for robust tNGS assay design.
- This review provides a comprehensive overview to accelerate diagnostic development.
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