title: A high-throughput Arabidopsis reverse genetics system.
abstract: A collection of Arabidopsis lines with T-DNA insertions in known sites was generated to increase the efficiency of functional genomics. A high-throughput modified thermal asymmetric interlaced (TAIL)-PCR protocol was developed and used to amplify DNA fragments flanking the T-DNA left borders from approximately 100000 transformed lines. A total of 85108 TAIL-PCR products from 52964 T-DNA lines were sequenced and compared with the Arabidopsis genome to determine the positions of T-DNAs in each line. Predicted T-DNA insertion sites, when mapped, showed a bias against predicted coding sequences. Predicted insertion mutations in genes of interest can be identified using Arabidopsis Gene Index name searches or by BLAST (Basic Local Alignment Search Tool) search. Insertions can be confirmed by simple PCR assays on individual lines. Predicted insertions were confirmed in 257 of 340 lines tested (76%). This resource has been named SAIL (Syngenta Arabidopsis Insertion Library) and is available to the scientific community at www.tmri.org.
authors.affiliation: Torrey Mesa Research Institute, Syngenta, 3115 Merryfield Row, San Diego, California 92121, USA. allen.sessions@syngenta.com
authors.forename: Allen
authors.initials: A
authors.lastname: Sessions
authors.forename: Ellen
authors.initials: E
authors.lastname: Burke
authors.forename: Gernot
authors.initials: G
authors.lastname: Presting
authors.forename: George
authors.initials: G
authors.lastname: Aux
authors.forename: John
authors.initials: J
authors.lastname: McElver
authors.forename: David
authors.initials: D
authors.lastname: Patton
authors.forename: Bob
authors.initials: B
authors.lastname: Dietrich
authors.forename: Patrick
authors.initials: P
authors.lastname: Ho
authors.forename: Johana
authors.initials: J
authors.lastname: Bacwaden
authors.forename: Cynthia
authors.initials: C
authors.lastname: Ko
authors.forename: Joseph D
authors.initials: JD
authors.lastname: Clarke
authors.forename: David
authors.initials: D
authors.lastname: Cotton
authors.forename: David
authors.initials: D
authors.lastname: Bullis
authors.forename: Jennifer
authors.initials: J
authors.lastname: Snell
authors.forename: Trini
authors.initials: T
authors.lastname: Miguel
authors.forename: Don
authors.initials: D
authors.lastname: Hutchison
authors.forename: Bill
authors.initials: B
authors.lastname: Kimmerly
authors.forename: Theresa
authors.initials: T
authors.lastname: Mitzel
authors.forename: Fumiaki
authors.initials: F
authors.lastname: Katagiri
authors.forename: Jane
authors.initials: J
authors.lastname: Glazebrook
authors.forename: Marc
authors.initials: M
authors.lastname: Law
authors.forename: Stephen A
authors.initials: SA
authors.lastname: Goff
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics; D017360:Arabidopsis / Q000235:genetics*; D001665:Binding Sites / Q000235:genetics; D032461:Chromosomes, Plant / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics*; D018744:DNA, Plant / Q000737:chemistry / Q000235:genetics; D030541:Databases, Genetic; D018745:Genome, Plant; D020407:Internet; D016254:Mutagenesis, Insertional; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction / Q000379:methods; D012639:Seeds / Q000235:genetics; D017422:Sequence Analysis, DNA
chemical_list: D004269:DNA, Bacterial; D018744:DNA, Plant; C036483:T-DNA