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Updated: Jan 6, 2026
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Detecting Somatic Genetic Alterations in Tumor Specimens by Exon Capture and Massively Parallel Sequencing
Published on: October 18, 2013
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Whole-Exome Sequencing (WES) for Illumina Short Read Sequencers Using Solution-Based Capture.
Milind C Mahajan1, Andrew S McLellan2
1Sema4, a Mount Sinai venture, Stamford, CT, USA.
Methods in Molecular Biology (Clifton, N.J.)
|October 6, 2019
Summary
Next-generation sequencing (NGS) and whole-exome sequencing (WES) advance genetic mutation identification for diagnostics and personalized medicine. This protocol details WES library preparation and exome enrichment for clinical applications.
Area of Science:
- Genomics
- Molecular Biology
- Clinical Diagnostics
Background:
- Next-generation sequencing (NGS) revolutionizes clinical research and diagnostics by identifying genetic mutations.
- Whole-exome sequencing (WES) targets coding regions, capturing most disease-causing mutations for comprehensive analysis.
- WES supports personalized medicine by informing diagnosis, therapeutic strategies, and disease risk prediction.
Purpose of the Study:
- To present a detailed protocol for performing whole-exome sequencing (WES).
- To outline steps for genomic DNA extraction, library preparation, and exome enrichment.
- To describe quality control methods including purification and qPCR for assessing capture efficiency.
Main Methods:
- Genomic DNA (gDNA) extraction from blood or saliva.
- Library preparation for Illumina sequencing instruments.
- Exome enrichment using Roche NimbleGen SeqCap EZ Exome v3 kit.
- Purification using AMPure XP System and quality assessment via qPCR.
Main Results:
- A standardized protocol for WES is established.
- Efficient purification and quality control methods are detailed.
- The protocol facilitates the identification of genetic variations for clinical applications.
Conclusions:
- The described WES protocol enables robust genetic analysis for clinical research and diagnostics.
- This method supports personalized medicine by identifying actionable genetic information.
- The protocol provides a foundation for utilizing WES data in understanding complex diseases.
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