title: Rapid, low-input, low-bias construction of shotgun fragment libraries by high-density in vitro transposition.
abstract: We characterize and extend a highly efficient method for constructing shotgun fragment libraries in which transposase catalyzes in vitro DNA fragmentation and adaptor incorporation simultaneously. We apply this method to sequencing a human genome and find that coverage biases are comparable to those of conventional protocols. We also extend its capabilities by developing protocols for sub-nanogram library construction, exome capture from 50 ng of input DNA, PCR-free and colony PCR library construction, and 96-plex sample indexing.
authors.affiliation: Department of Genome Sciences, University of Washington, Seattle, WA 98195, USA. acadey@uw.edu
authors.forename: Andrew
authors.initials: A
authors.lastname: Adey
authors.forename: Hilary G
authors.initials: HG
authors.lastname: Morrison
authors.lastname: Asan
authors.forename: Xu
authors.initials: X
authors.lastname: Xun
authors.forename: Jacob O
authors.initials: JO
authors.lastname: Kitzman
authors.forename: Emily H
authors.initials: EH
authors.lastname: Turner
authors.forename: Bethany
authors.initials: B
authors.lastname: Stackhouse
authors.forename: Alexandra P
authors.initials: AP
authors.lastname: MacKenzie
authors.forename: Nicholas C
authors.initials: NC
authors.lastname: Caruccio
authors.forename: Xiuqing
authors.initials: X
authors.lastname: Zhang
authors.forename: Jay
authors.initials: J
authors.lastname: Shendure
mesh_terms: D000818:Animals; D004247:DNA / Q000302:isolation & purification; D053938:DNA Fragmentation; D004330:Drosophila / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005091:Exons; D015894:Genome, Human; D049750:Genome, Insect; D015698:Genomic Library; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA / Q000379:methods*; D019895:Transposases / Q000378:metabolism
chemical_list: D004247:DNA; D019895:Transposases