Protocol for rapid allelic discrimination qPCR genotyping of the Winnie mouse model.

Winnie mice are a widely used in vivo model of inflammatory bowel disease and carry a missense mutation in the Muc2 gene. Here, we present a protocol for genotyping Winnie mice using TaqMan allelic discrimination quantitative PCR. We describe tissue collection, rapid crude DNA extraction, probe-based amplification with dual-labeled fluorophores, and fluorescence-based genotype calling in a single reaction. This protocol enables qualitative SNP genotyping without post-amplification processing and can be readily adapted to other defined point mutations.