SNP Genotyping with Amplifluor-Like Method
Khassanova G. Khalbayeva S. Serikbay D. Mazkirat S. Bulatova K. Utebayev M. Shavrukov Y.
2023NLM (Medline)
Methods in molecular biology (Clifton, N.J.)
2023#2638201 - 219 pp.
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.
Amplifluor-like , Fluorescence , FRET , Genotyping , SNP , Universal molecular probes
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A.I. Barayev Research and Production Centre of Grain Farming, Shortandy, Kazakhstan
Faculty of Agronomy, S. Seifullin Kazakh AgroTechnical University, Astana, Kazakhstan
Kazakh Research Institute of Agriculture and Plant Production, Almaty, Kazakhstan
College of Agronomy, Northwest A&F University, Yangling, Shaanxi, China
College of Science and Engineering, Biological Sciences, Flinders University, Adelaide, SA, Australia
A.I. Barayev Research and Production Centre of Grain Farming
Faculty of Agronomy
Kazakh Research Institute of Agriculture and Plant Production
College of Agronomy
College of Science and Engineering
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