Torres Anthony, Argyris Jason, Gaudino Reginald
Cannabis Research Institute, Discovery Partners Institute, University of Illinois System, Chicago, IL, USA.
Centre for Research in Agricultural Genomics (CRAG), CSIC-IRTA-UAB-UB, Campus, UAB, Barcelona, Spain.
Methods Mol Biol. 2025;2943:143-155. doi: 10.1007/978-1-0716-4642-7_12.
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.
本章介绍了植物科学中先进的基因分型和遗传分析方案，尤其侧重于利用从FTA样本和叶片组织中提取的DNA。PACE（PCR等位基因竞争性延伸）能够以极高的准确性实现高通量SNP基因分型，特别针对并区分两个感兴趣的等位基因。该方案涉及使用等位基因特异性引物和PACE预混液进行选择性延伸和标记，最终清晰地鉴定SNP基因型。此方法为SNP基因分型提供了一个有价值的工具，可有效检测与所需性状相关的特定且靶向的遗传变异。在此，我们描述了一种简化的工作流程，用于使用此方法对雌雄异株植物的目标基因中的SNP进行基因分型。
