Identification of the oim mutation by dye terminator chemistry combined with automated direct DNA sequencing

N. P. Camacho, D. Dow, T. R. Toledano, J. K. Buckmeyer, J. M. Gertner, Cory Brayton, C. L. Raggio, L. Root, A. L. Boskey

Research output: Contribution to journalArticle

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 proα2(I) collagen gene) that results in accumulation of α1(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.

Original languageEnglish (US)
Pages (from-to)38-42
Number of pages5
JournalJournal of Orthopaedic Research
Volume16
Issue number1
DOIs
StatePublished - Jan 1998
Externally publishedYes

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DNA Sequence Analysis
Coloring Agents
Mutation
Collagen
Osteogenesis Imperfecta
Sequence Deletion
Heterozygote
Genes
Nucleotides
Genotype
Phenotype

ASJC Scopus subject areas

  • Orthopedics and Sports Medicine

Cite this

Identification of the oim mutation by dye terminator chemistry combined with automated direct DNA sequencing. / Camacho, N. P.; Dow, D.; Toledano, T. R.; Buckmeyer, J. K.; Gertner, J. M.; Brayton, Cory; Raggio, C. L.; Root, L.; Boskey, A. L.

In: Journal of Orthopaedic Research, Vol. 16, No. 1, 01.1998, p. 38-42.

Research output: Contribution to journalArticle

Camacho, NP, Dow, D, Toledano, TR, Buckmeyer, JK, Gertner, JM, Brayton, C, Raggio, CL, Root, L & Boskey, AL 1998, 'Identification of the oim mutation by dye terminator chemistry combined with automated direct DNA sequencing', Journal of Orthopaedic Research, vol. 16, no. 1, pp. 38-42. https://doi.org/10.1002/jor.1100160107
Camacho, N. P. ; Dow, D. ; Toledano, T. R. ; Buckmeyer, J. K. ; Gertner, J. M. ; Brayton, Cory ; Raggio, C. L. ; Root, L. ; Boskey, A. L. / Identification of the oim mutation by dye terminator chemistry combined with automated direct DNA sequencing. In: Journal of Orthopaedic Research. 1998 ; Vol. 16, No. 1. pp. 38-42.
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