title: Identification of the oim mutation by dye terminator chemistry combined with automated direct DNA sequencing.
abstract: The homozygous oim/oim mouse, a model of moderate-to-severe human osteogenesis imperfecta, contains a G-nucleotide deletion in the Cola-2 gene (the murine pro alpha(I) collagen gene) that results in accumulation of alpha1(I) homotrimer collagen. Although these mice have a distinctive phenotype that includes multiple fractures and deformities, genotyping is necessary to distinguish them from their wildtype (+/+) and heterozygote (oim/+) littermates. In this study, the dye primer and dye terminator chemistry methods, in combination with automated direct DNA sequencing, were compared for accuracy and ease in genotyping. A total of 82 mice from 14 litters were bred and genotyped; this resulted in 18 +/+, 35 oim/+, and 29 oim/oim mice. The dye primer and dye terminator chemistry methods worked equally well for identification of the deletion mutation and thus the genotype of all of the mice. However, the dye terminator method was found to be superior on the basis of the reduced amount of sample handling and reduced quantity of reagent required.
authors.affiliation: Research Division, The Hospital for Special Surgery, New York, New York 10021, USA. camachon@hss.edu
authors.forename: N P
authors.initials: NP
authors.lastname: Camacho
authors.forename: D
authors.initials: D
authors.lastname: Dow
authors.forename: T R
authors.initials: TR
authors.lastname: Toledano
authors.forename: J K
authors.initials: JK
authors.lastname: Buckmeyer
authors.forename: J M
authors.initials: JM
authors.lastname: Gertner
authors.forename: C F
authors.initials: CF
authors.lastname: Brayton
authors.forename: C L
authors.initials: CL
authors.lastname: Raggio
authors.forename: L
authors.initials: L
authors.lastname: Root
authors.forename: A L
authors.initials: AL
authors.lastname: Boskey
mesh_terms: D000818:Animals; D003094:Collagen / Q000235:genetics*; D004396:Coloring Agents; D005838:Genotype; D051379:Mice; D009154:Mutation; D010013:Osteogenesis Imperfecta / Q000235:genetics*; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
chemical_list: D004396:Coloring Agents; D003094:Collagen