A.I. Barayev Research and Production Centre of Grain Farming, Shortandy, Kazakhstan.
Faculty of Agronomy, S. Seifullin Kazakh AgroTechnical University, Astana, Kazakhstan.
Methods Mol Biol. 2023;2638:201-219. doi: 10.1007/978-1-0716-3024-2_14.
For SNP genotyping, amplification of fluorescence (Amplifluor) is a popular and actively developing method in the plant sciences. The "Amplifluor-like" is a "home-made" modification of the original commercial Amplifluor method. Amplifluor-like genotyping requires two essential components: (1) two allele-specific forward primers targeting the SNP site with one common reverse primer; and (2) a universal part with two non-allele-specific molecular probes containing one of the two used fluorophores and a quencher. Allele discrimination is based on the fluorescence score, where the dominance of one dye over the other confirms the presence of each specific SNP allele. The Amplifluor-like method is similar to commercial KASP and original Amplifluor methods but is much cheaper because all components can be ordered as regular and modified oligos. The easily adaptable Amplifluor-like method can be modified by any researcher to make it suitable for available instruments, reagents and conditions in low-budget laboratories for SNP genotyping of any plant species with identified genetic polymorphism.
对于 SNP 基因分型，荧光扩增（Amplifluor）是植物科学中一种流行且正在积极发展的方法。“Amplifluor 类似物”是对原始商业 Amplifluor 方法的“自制”修改。Amplifluor 类似物基因分型需要两个基本组件：（1）两个针对 SNP 位点的等位基因特异性正向引物，一个共同的反向引物；（2）一个通用部分，包含两个非等位基因特异性分子探针，其中一个含有两种荧光染料之一和一个淬灭剂。等位基因的区分基于荧光评分，其中一种染料对另一种染料的优势证实了每种特定 SNP 等位基因的存在。Amplifluor 类似物方法类似于商业的 KASP 和原始的 Amplifluor 方法，但价格便宜得多，因为所有组件都可以作为常规和修饰的寡核苷酸订购。这种易于适应的 Amplifluor 类似物方法可以由任何研究人员进行修改，以使其适用于低成本实验室中可用的仪器、试剂和条件，用于具有已确定遗传多态性的任何植物物种的 SNP 基因分型。
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