Acadl-SNP based genotyping assay for long-chain acyl-CoA dehydrogenase deficient mice

The long-chain acyl-CoA dehydrogenase (LCAD) (Acadl = gene; LCAD = protein) deficient mouse model has been important in evaluating the role of mitochondrial fatty acid oxidation of long-chain fatty acids in metabolic disorders. The insertion vector-based gene targeting strategy used to generate this model has made it difficult to distinguish homozygous and heterozygous genotypes containing targeted Acadl alleles in LCAD-deficient mice. Herein, we describe the design and validation of Acadl SNP genotyping methods capable of distinguishing between heterozygous and homozygous LCAD-deficient mice. The Acadl SNP genotyping assays are effective at allelic discrimination of both C57BL/6 and 129 mouse strain-based Acadl alleles under conditions including, both low purity and quantity genomic DNA templates. This makes the method practical and provides the necessary tools for genotyping the LCAD-deficient mouse model.

► Genotyping of Acadl targeted alleles in LCAD-deficient mice has been impractical. ► We have developed Acadl SNP genotyping assays for LCAD-deficient mice. ► The Acadl SNP genotyping assays are effective at both low DNA purity and quantity.