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Updated: Apr 22, 2026
Targeted DNA Methylation Analysis by Next-generation Sequencing
Published on: February 24, 2015
Highly sensitive targeted methylome sequencing by post-bisulfite adaptor tagging.
1Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST), Higashi-ku, Fukuoka 812-8582, Japan Department of Biochemistry, Kyushu University Graduate School of Medical Sciences, Higashi-ku, Fukuoka 812-8582, Japan.
Targeted methylome sequencing (TMS) is now more accessible for limited DNA samples. Post-bisulfite adaptor tagging (PBAT) enables cost-effective epigenomic analysis with 100x less input DNA.
Area of Science:
- Epigenetics
- Genomics
- Molecular Biology
Background:
- Whole-genome bisulfite sequencing (WGBS) is the gold standard for methylome analysis but is costly for large-scale studies.
- Targeted methylome sequencing (TMS) offers a cost-effective alternative but requires substantial input DNA.
- Current TMS protocols are unsuitable for precious samples with limited DNA quantity.
Purpose of the Study:
- To develop a more accessible and cost-effective method for targeted methylome sequencing.
- To enable methylome analysis using significantly reduced amounts of input DNA.
- To overcome limitations of existing TMS protocols for epigenomic research.
Main Methods:
- Developed a novel method using post-bisulfite adaptor tagging (PBAT).
- PBAT involves adaptor tagging after bisulfite treatment to preserve DNA templates.
- Reduced input DNA requirements by 100-fold and decreased PCR cycles compared to standard TMS.
Main Results:
- PBAT enables targeted methylome sequencing (TMS) with significantly less input DNA.
- The method circumvents DNA template loss during bisulfite treatment.
- Achieved substantial reduction in DNA input and PCR amplification cycles.
Conclusions:
- PBAT-mediated TMS is a flexible and cost-effective epigenomic analysis tool.
- This method significantly expands the applicability of TMS to samples with limited DNA.
- PBAT-mediated TMS is expected to become an invaluable technique in epigenomics research.
