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Single-Base Extension and ELISA-Based Approach for Single-Nucleotide Polymorphisms Genotyping
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Single-Base Extension and ELISA-Based Approach for Single-Nucleotide Polymorphisms Genotyping
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Journal Article
Single-Base Extension and ELISA-Based Approach for Single-Nucleotide Polymorphisms Genotyping
2011
Overview
Single-nucleotide polymorphisms (SNPs) emerge as a fundamental tool in personalized medicine due to their association with drug responses or disease predisposition. Single-base extension (SBE) is a common method for characterizing known SNPs, but involves complicated procedures or requires costly analytical instruments. Here, we describe a novel SNP genotyping based on SBE and enzyme-linked immunosorbent assay (ELISA). During the SBE, the 5′ end fluorescein isothiocyanate-labeled allele-specific primer will extend with biotinylated dideoxynucleotides which are complementary to the SNP sites. The extension product will then be captured by streptavidin-coated nanoparticle and develop blue color in the ELISA assay. We validated this method by detecting SNPs for TP53 gene codon 273 from 68 individuals and the data were 100% in concordant with DNA sequencing. Thus, SBE and ELISA-based SNPs assay is a simple and accurate method for SNP genotyping.
Publisher
New York : Humana Press Inc,Humana Press Inc,Springer,Springer Nature B.V
Subject
/ Biological and medical sciences
/ Chemistry and Materials Science
/ color
/ drugs
/ enzyme-linked immunosorbent assay
/ Enzyme-Linked Immunosorbent Assay - methods
/ Enzymes
/ Fundamental and applied biological sciences. Psychology
/ genes
/ genetics
/ Genotype
/ Humans
/ medicine
/ methods
/ Polymorphism, Single Nucleotide
