Single-nucleotide polymorphism analysis using fluorescence resonance energy transfer between DNA-labeling fluorophore, fluorescein isothiocyanate, and DNA intercalator, POPO-3, on bacterial magnetic particles

Hideki Nakayama, Atsushi Arakaki, Kohei Maruyama, Haruko Takeyama, Tadashi Matsunaga

Research output: Contribution to journalArticle

54 Citations (Scopus)

Abstract

To develop an analytical system for single-nucleotide polymorphisms (SNPs), the fluorescence resonance energy transfer (FRET) technique was employed on a bacterial magnetic particle (BMP) surface. A combination of fluorescein isothiocyanate (FITC; excitation 490 nm/emission 520 nm) labeled at the 5′ end of DNA and an intercalating compound (POPO-3, excitation 534 nm/emission 570 nm) was used to avoid the interference from light scattering caused by nanoparticles. After hybridization between target DNA immobilized onto BMPs and FITC-labeled probes, fluorescence from POPO-3, which was excited by the energy from the FITC, was detected. The major homozygous (ALDH2*1), heterozygous (ALDH2*1/*2), and minor homozygous (ALDH2*2) genotypes in the blood samples were discriminated by this method. The assay described herein allows for a simple and rapid SNP analysis using a fully automated system.

Original languageEnglish
Pages (from-to)96-102
Number of pages7
JournalBiotechnology and bioengineering
Volume84
Issue number1
DOIs
Publication statusPublished - 2003 Oct 5
Externally publishedYes

Keywords

  • ALDH2
  • Bacterial magnetic particles (BMPs)
  • Flourescence resonance energy transfer (FRET)
  • Hybridization
  • Single nucleotide polymorphism (SNP) analysis

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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