Protocol for rapid allelic discrimination qPCR genotyping of the Winnie mouse model
Year: 2026
Type: article
Abstract: 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. Highlights: 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.
Source: STAR Protocols
Authors: Behzad Mansoori, Chengyu Liang
Institution: The Wistar Institute
Language: English
Topic: Inflammatory Bowel Disease
Subfield: Genetics
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $1,550
OpenAlex ID: W7153585134