Torres Anthony, 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;2943:177-188. doi: 10.1007/978-1-0716-4642-7_15.
This chapter presents protocols for advanced genotyping and genetic analysis in plant science, utilizing DNA extracted from FTA samples and leaf tissue. Targeted long range PCR is employed to uncover allelic and structural variations within genes of interest, while targeted long-read nanopore sequencing offers a comprehensive view of gene targets in their native and enriched forms. The protocol for nanopore sequencing encompasses DNA preparation, barcoding of up to 96 samples, adapter ligation, purification, and sequencing, culminating in the generation of raw data files (pod5 or fastq) suitable for further analysis. Together, these methods provide a powerful toolkit for plant diagnostics and breeding.
本章介绍了植物科学中先进的基因分型和遗传分析方案，利用从FTA样本和叶片组织中提取的DNA。靶向长程PCR用于揭示感兴趣基因内的等位基因和结构变异，而靶向长读长纳米孔测序则能全面呈现目标基因的天然和富集形式。纳米孔测序方案包括DNA制备、多达96个样本的条形码标记、接头连接、纯化和测序，最终生成适合进一步分析的原始数据文件（pod5或fastq）。这些方法共同为植物诊断和育种提供了一个强大的工具包。
