PCR Allele Competitive Extension (PACE) Workflow for Genotyping by SNP Identification

This chapter presents protocols for advanced genotyping and genetic analysis in plant science, particularly utilizing DNA extracted from FTA samples and leaf tissue. PACE (PCR Allele Competitive Extension) enables high-throughput SNP genotyping with remarkable accuracy, specifically targeting and differentiating between two alleles of interest. The protocol involves using allele-specific primers and a PACE master mix for selective extension and labeling, culminating in the clear identification of SNP genotypes. This method provides a valuable tool for SNP genotyping, efficiently detecting specific and targeted genetic variations associated with desired traits. Here, we describe a simplified workflow for SNP genotyping using this approach to detect SNPs in target genes of a dioecious plant.