title: Ultra-low-input, tagmentation-based whole-genome bisulfite sequencing.
abstract: We have adapted transposase-based in vitro shotgun library construction ("tagmentation") for whole-genome bisulfite sequencing. This method, Tn5mC-seq, enables a >100-fold reduction in starting material relative to conventional protocols, such that we generate highly complex bisulfite sequencing libraries from as little as 10 ng of input DNA, and ample useful sequences from 1 ng of input DNA. We demonstrate Tn5mC-seq by sequencing the methylome of a human lymphoblastoid cell line to ∼8.6× high-quality coverage of each strand.
authors.affiliation: Department of Genome Sciences, University of Washington, Seattle, WA 98195, USA.
authors.forename: Andrew
authors.initials: A
authors.lastname: Adey
authors.forename: Jay
authors.initials: J
authors.lastname: Shendure
mesh_terms: D001483:Base Sequence; D002461:Cell Line, Transformed; D018899:CpG Islands; D003596:Cytosine / Q000737:chemistry; D004247:DNA / Q000187:drug effects; D019175:DNA Methylation; D015723:Gene Library; D015894:Genome, Human; D006801:Humans; D008214:Lymphocytes / Q000473:pathology / Q000502:physiology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA / Q000379:methods*; D013447:Sulfites / Q000494:pharmacology; D019895:Transposases / Q000235:genetics
chemical_list: D013447:Sulfites; D003596:Cytosine; D004247:DNA; D019895:Transposases; C042345:hydrogen sulfite