期刊
DNA RESEARCH
卷 22, 期 1, 页码 13-18出版社
OXFORD UNIV PRESS
DOI: 10.1093/dnares/dsu034
关键词
DNA methylation; target enrichment; massively parallel sequencing
资金
- Research Program of Innovative Cell Biology by Innovative Technology (Cell Innovation)
- Platform for Drug Discovery, Informatics and Structural Life Science
- Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan [25129702]
- MEXT
The current gold standard method for methylome analysis is whole-genome bisulfite sequencing (WGBS), but its cost is substantial, especially for the purpose of multi-sample comparison of large methylomes. Shotgun bisulfite sequencing of target-enriched DNA, or targeted methylome sequencing (TMS), can be a flexible, cost-effective alternative to WGBS. However, the current TMS protocol requires a considerable amount of input DNA and hence is hardly applicable to samples of limited quantity. Here we report a method to overcome this limitation by using post-bisulfite adaptor tagging (PBAT), in which adaptor tagging is conducted after bisulfite treatment to circumvent bisulfite-induced loss of intact sequencing templates, thereby enabling TMS of a 100-fold smaller amount of input DNA with far fewer cycles of polymerase chain reaction than in the current protocol. We thus expect that the PBAT-mediated TMS will serve as an invaluable method in epigenomics.
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