title: Mu-seq: sequence-based mapping and identification of transposon induced mutations.
abstract: Mutations tagged by transposon insertions can be readily mapped and identified in organisms with sequenced genomes. Collections of such mutants allow a systematic analysis of gene function, and can be sequence-indexed to build invaluable resources. Here we present Mu-seq (Mutant-seq), a high-throughput NextGen sequencing method for harnessing high-copy transposons. We illustrate the efficacy of Mu-seq by applying it to the Robertson's Mutator system in a large population of maize plants. A single Mu-seq library, for example, constructed from 576 different families (2304 plants), enabled 4, 723 novel, germinal, transposon insertions to be detected, identified, and mapped with single base-pair resolution. In addition to the specificity, efficiency, and reproducibility of Mu-seq, a key feature of this method is its adjustable scale that can accommodate simultaneous profiling of transposons in thousands of individuals. We also describe a Mu-seq bioinformatics framework tailored to high-throughput, genome-wide, and population-wide analysis of transposon insertions.
authors.affiliation: Horticultural Sciences Department, Plant Molecular and Cellular Biology Program, Genetics Institute, University of Florida, Gainesville, Florida, United States of America.
authors.forename: Donald R
authors.initials: DR
authors.lastname: McCarty
authors.forename: Sue
authors.initials: S
authors.lastname: Latshaw
authors.forename: Shan
authors.initials: S
authors.lastname: Wu
authors.forename: Masaharu
authors.initials: M
authors.lastname: Suzuki
authors.forename: Charles T
authors.initials: CT
authors.lastname: Hunter
authors.forename: Wayne T
authors.initials: WT
authors.lastname: Avigne
authors.forename: Karen E
authors.initials: KE
authors.lastname: Koch
mesh_terms: D001483:Base Sequence; D019295:Computational Biology / Q000379:methods*; D004251:DNA Transposable Elements / Q000235:genetics*; D015723:Gene Library; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics*; D009841:Oligonucleotides / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods
chemical_list: D004251:DNA Transposable Elements; D009841:Oligonucleotides