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 carrying 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.

Allelic discrimination qPCR protocol for genotyping the Muc2 p.Cys52Tyr mutation using dual-labeled hydrolysis probes Enables rapid discrimination of wild-type, heterozygous, and mutant alleles in a single reactionCompatible with standard real-time PCR instruments and requires no post-PCR processingSupports high-throughput genotyping from crude DNA with minimal hands-on time.