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POSTER 91 - USE OF CDNA ARRAY HYBRIDIZATION AND EXPRESSION ANALYSES IN GENE TESTING FOR ENU-INDUCED MUTATIONS IN MICE
P Charmley
Celltech R&D,
Inc.
Bennington A,
Snell A, Appleby M, Strobel S
Celltech R&D,
Inc
As part of a mutant gene testing approach in our ENU mouse mutagenesis program, we are using oligo-based array hybridization technology to aid in finding the ENU-induced phenotype-causing mutation in our mutant mice. Our application of this technology has involved the duplicate spotting of 60-mer oligos from every known or predicted gene in the genetically mapped interval that contains the phenotype-causing mutation. We have used real-time PCR to assess the robustness of the array technology through side-by-side comparisons of the expression measurements. By our first applying this array hybridization approach to an interval with a known ENU mutation we are gaining confidence in the strengths and limits of this technology. We are using the expression results to prioritize which genes are sequenced for the mutation, as well as the possibility for identifying mutants that alter the gene expression levels. We hope our practical experiences in using gene expression array technology are helpful to other groups who plan to use this approach to gene expression measurements in pursuit of ENU-induced mutations in mice.
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