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Background  Mice carrying the spontaneous genetic mutation known as Wallerian degeneration slow ( Wld   s  ) have a unique neuroprotective phenotype, where axonal and synaptic compartments of neurons are protected from degeneration following a wide variety of physical, toxic and inherited disease-inducing stimuli. This remarkable phenotype has been shown to delay onset and progression in several mouse models of neurodegenerative disease, suggesting that  Wld   s  -mediated neuroprotection may assist in the identification of novel therapeutic targets. As a result, cross-breeding of  Wld   s   mice with mouse models of neurodegenerative diseases is used increasingly to understand the roles of axon and synapse degeneration in disease. However, the phenotype shows strong gene-dose dependence so it is important to distinguish offspring that are homozygous or heterozygous for the mutation. Since the  Wld   s   mutation comprises a triplication of a region already present in the mouse genome, the most stringent way to quantify the number of mutant  Wld   s   alleles is using copy number. Current approaches to genotype  Wld   s   mice are based on either Southern blots or pulsed field gel electrophoresis, neither of which are as rapid or efficient as quantitative PCR (QPCR).    Results  We have developed a rapid, robust and efficient genotyping method for  Wld   s   using QPCR. This approach differentiates, based on copy number, homozygous and heterozygous  Wld   s   mice from wild-type mice and each other. We show that this approach can be used to genotype mice carrying the spontaneous  Wld   s   mutation as well as animals expressing the  Wld   s   transgene.    Conclusion  We have developed a QPCR genotyping method that permits rapid and effective genotyping of  Wld   s   copy number. This technique will be of particular benefit in studies where  Wld   s   mice are cross-bred with other mouse models of neurodegenerative disease in order to understand the neuroprotective processes conferred by the  Wld   s   mutation.

Design of a novel quantitative PCR (QPCR)-based protocol for genotyping mice carrying the neuroprotective Wallerian degeneration slow (Wlds) gene