title: Highly sensitive targeted methylome sequencing by post-bisulfite adaptor tagging.
abstract: 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.
authors.affiliation: Department 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.
authors.forename: Fumihito
authors.initials: F
authors.lastname: Miura
authors.affiliation: Department 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 tito@med.kyushu-u.ac.jp.
authors.forename: Takashi
authors.initials: T
authors.lastname: Ito
mesh_terms: D000818:Animals; D002460:Cell Line; D019175:DNA Methylation / Q000502:physiology*; D044127:Epigenesis, Genetic / Q000502:physiology*; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D051379:Mice; D013447:Sulfites / Q000737:chemistry
chemical_list: D013447:Sulfites; C042345:hydrogen sulfite
keywords: DNA methylation; massively parallel sequencing; target enrichment