Torres Anthony, Givens Robert, Gaudino Reginald
Cannabis Research Institute, Discovery Partners Institute, University of Illinois System, Chicago, IL, USA.
Front Range Biosciences, Lafayette, CO, USA.
Methods Mol Biol. 2025;2943:131-141. doi: 10.1007/978-1-0716-4642-7_11.
This chapter outlines a basic protocol for DNA extraction, applicable to both FTA cards and leaf pieces. The extracted DNA then undergoes a pre-amplification enrichment reaction, followed by dilution and preparation for input into a real-time amplification with High-Resolution Melt (HRM) detection. The resulting melting genotypes are produced for comparison and analysis, providing a practical approach for genotyping studies. The flexibility of the method allows specific analytical output and high throughput analysis of a given DNA sequence that possesses inherent variability between individuals. This protocol describes the crude approach of extracting DNA from samples, preparing a real-time HRM reaction, and analyzing the result.
本章概述了一种适用于FTA卡和叶片片段的DNA提取基本方案。提取的DNA随后进行预扩增富集反应，接着稀释并准备用于高分辨率熔解（HRM）检测的实时扩增。生成所得的熔解基因型用于比较和分析，为基因分型研究提供了一种实用方法。该方法的灵活性允许对具有个体间固有变异性的给定DNA序列进行特定的分析输出和高通量分析。本方案描述了从样品中提取DNA、制备实时HRM反应以及分析结果的粗略方法。
