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

Fingerprint

Intercalating Agents
Fluorescence Resonance Energy Transfer
Fluorophores
Fluorescein-5-isothiocyanate
Nucleotides
Polymorphism
Fluorescein
Labeling
Single Nucleotide Polymorphism
DNA
Immobilized Nucleic Acids
Light scattering
Assays
Blood
Fluorescence
Nanoparticles
Genotype
Light
POPO 3
isothiocyanic acid

Keywords

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

ASJC Scopus subject areas

  • Biotechnology
  • Microbiology

Cite this

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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.",
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AU - Nakayama, Hideki

AU - Arakaki, Atsushi

AU - Maruyama, Kohei

AU - Takeyama, Haruko

AU - Matsunaga, Tadashi

PY - 2003/10/5

Y1 - 2003/10/5

N2 - 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.

AB - 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.

KW - ALDH2

KW - Bacterial magnetic particles (BMPs)

KW - Flourescence resonance energy transfer (FRET)

KW - Hybridization

KW - Single nucleotide polymorphism (SNP) analysis

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