Abstract
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.
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Acknowledgments
We thank Dr. Yingqun Wang for suggestion and editorial assistance. This work was supported by funding from Science and Technology Bureau of Heilongjiang Province (grant no. LC06C25).
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The authors declare no competing interests.
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Liu, G., Cheng, Y., Zhao, W. et al. Single-Base Extension and ELISA-Based Approach for Single-Nucleotide Polymorphisms Genotyping. Appl Biochem Biotechnol 163, 573–576 (2011). https://doi.org/10.1007/s12010-010-9063-4
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DOI: https://doi.org/10.1007/s12010-010-9063-4