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Targeted DNA Methylation Analysis by Next-generation Sequencing
Published on: February 24, 2015
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HAM-TBS: high-accuracy methylation measurements via targeted bisulfite sequencing
Simone Roeh1, Tobias Wiechmann1, Susann Sauer1
1Department of Translational Research in Psychiatry, Max Planck Institute of Psychiatry, Kraepelinstr. 2-10, 80804, Munich, Germany.
Epigenetics & Chromatin
|July 6, 2018
Summary
We developed HAM-TBS, a highly accurate and cost-effective bisulfite sequencing method to measure DNA methylation. This technique enables precise analysis of epigenetic modifications in multiple samples for disease research.
Area of Science:
- Epigenetics
- Molecular Biology
- Genomics
Background:
- Accurate measurement of DNA methylation is crucial for understanding epigenetic mechanisms and their role in common diseases.
- DNA methylation patterns are increasingly recognized as key players in the pathophysiology of various disorders.
- Existing methods may lack the accuracy, efficiency, or cost-effectiveness required for comprehensive methylation analysis.
Purpose of the Study:
- To present a novel, highly accurate method for measuring DNA methylation using bisulfite sequencing, termed HAM-TBS.
- To enable cost-effective, high-accuracy DNA methylation assessment across multiple samples.
- To develop and validate a targeted assay for the FKBP5 locus, relevant to stress-related disorders.
Main Methods:
- Developed HAM-TBS (Highly Accurate Methylation via Targeted Bisulfite Sequencing).
- Incorporated triplicate bisulfite conversion, pooled target enrichment, PCR-free library prep, and high-coverage sequencing (≥1000×).
- Enabled multiplexed analysis of up to 96 samples and 10 kb regions using Illumina MiSeq.
Main Results:
- HAM-TBS achieved a mean accuracy of 0.72% in resolving DNA methylation levels.
- The method demonstrated high accuracy and cost-effectiveness for multiplexed sample analysis.
- A targeted panel for the FKBP5 locus was designed and validated, covering regions not available on commercial arrays.
Conclusions:
- HAM-TBS is a robust, medium-throughput sequencing approach for precise DNA methylation detection.
- This method facilitates accurate assessment of methylation changes in specific genomic regions.
- HAM-TBS offers a valuable tool for advancing epigenetic research in disease contexts.
Keywords:
5-methylcytosineDNA methylationFKBP5Next-generation sequencingTargeted bisulfite sequencingMore Related Videos
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