SNP Genotyping with Amplifluor-Like Method

Gulmira Khassanova, Sholpan Khalbayeva, Dauren Serikbay, Shynar Mazkirat, Kulpash Bulatova, Maral Utebayev, Yuri Shavrukov

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review


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.

Original languageEnglish
Title of host publicationPlant Genotyping
Subtitle of host publicationMethods and Protocols
EditorsYuri Shavrukov
Place of PublicationNew York
PublisherSpringer Nature
Number of pages19
ISBN (Electronic)9781071630242
ISBN (Print)9781071630235
Publication statusPublished - 2023

Publication series

NameMethods in molecular biology (Clifton, N.J.)
PublisherSpringer Nature
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029


  • Amplifluor-like
  • Fluorescence
  • FRET
  • Genotyping
  • SNP
  • Universal molecular probes


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