Amplifluor-Based SNP Genotyping

Manmode Darpan Mohanrao; Senapathy Senthilvel; Yarabapani Rushwanth Reddy; Chippa Anil Kumar; Palchamy Kadirvel

doi:10.1007/978-1-0716-3024-2_13

Amplifluor-Based SNP Genotyping

Methods Mol Biol. 2023:2638:191-200. doi: 10.1007/978-1-0716-3024-2_13.

Authors

Manmode Darpan Mohanrao¹, Senapathy Senthilvel¹, Yarabapani Rushwanth Reddy¹, Chippa Anil Kumar¹, Palchamy Kadirvel²

Affiliations

¹ ICAR-Indian Institute of Oilseeds Research, Hyderabad, India.
² ICAR-Indian Institute of Oilseeds Research, Hyderabad, India. [email protected].

PMID: 36781643
DOI: 10.1007/978-1-0716-3024-2_13

Abstract

Amplifluor, a genotyping system used to analyze single nucleotide polymorphisms (SNPs), is supplied by Merck-Millipore. Amplifluor is based on polymerase chain reaction (PCR) with two competing allele-specific primers and a SNP specific common reverse primer. Sequence information flanking SNP of interest and fluorescent plate reader for end-point measurement or qPCR machine for real time measurement are required for the execution of the Amplifluor assay. In this chapter, the principle and working protocol of the Amplifluor assay based on end-point fluorescence detection of SNP allele is presented with an example.

Keywords: Amplifluor; Genotypic assays; Single nucleotide polymorphisms (SNPs).

MeSH terms

Alleles
DNA Primers / genetics
Genotype
Polymerase Chain Reaction / methods
Polymorphism, Single Nucleotide*

Substances

DNA Primers