Development and evaluation of an automated workstation for single nucleotide polymorphism discrimination using bacterial magnetic particles

Tsuyoshi Tanaka, Kohei Maruyama, Kiyoushi Yoda, Etsuo Nemoto, Yuuji Udagawa, Hideki Nakayama, Haruko Takeyama, Tadashi Matsunaga

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

We designed an automated workstation for magnetic particle-based single nucleotide polymorphism (SNP) discrimination of ALDH genotypes. Bacterial magnetic particles (BMPs) extracted from Magnetospirillum magneticum AMB-1 were used as DNA carriers. The principle for SNP discrimination in this study was based on fluorescence resonance energy transfer (FRET) between FITC (donor) and POPO-3 (acceptor) bound to double-stranded DNA. The workstation is equipped with a 96-way automated pipetter which collects and dispenses fluids as it moves in x- and z-directions. The platform contains a disposable tip rack station, a reagent vessel serving as a stock for POPO-3 and FITC-labeled probes and a reaction station for a 96-well microtiter plate. BMPs were collected by attaching a neodymium iron boron sintered (Nd-Fe-B) magnet on the bottom of the microtiter plate. This system permits the simultaneous heating and magnetic separation of 96 samples per assay. The genotypes ALDH2*1 and ALDH2*2 were discriminated by calculating the relative fluorescence intensities on BMPs.

Original languageEnglish
Pages (from-to)325-330
Number of pages6
JournalBiosensors and Bioelectronics
Volume19
Issue number4
DOIs
Publication statusPublished - 2003 Dec 15
Externally publishedYes

Fingerprint

Fluorescein-5-isothiocyanate
Nucleotides
Polymorphism
Single Nucleotide Polymorphism
Magnetospirillum
DNA
Genotype
Neodymium
Magnetic separation
Fluorescence Resonance Energy Transfer
Boron
Magnets
Heating
Assays
Iron
Fluorescence
Fluids
POPO 3
Direction compound

Keywords

  • Aldehyde dehydrogenase 2 gene
  • Bacterial magnetic particles (BMPs)
  • Fluorescence resonance energy transfer
  • Single nucleotide polymorphism (SNP)

ASJC Scopus subject areas

  • Biotechnology
  • Analytical Chemistry
  • Electrochemistry

Cite this

Development and evaluation of an automated workstation for single nucleotide polymorphism discrimination using bacterial magnetic particles. / Tanaka, Tsuyoshi; Maruyama, Kohei; Yoda, Kiyoushi; Nemoto, Etsuo; Udagawa, Yuuji; Nakayama, Hideki; Takeyama, Haruko; Matsunaga, Tadashi.

In: Biosensors and Bioelectronics, Vol. 19, No. 4, 15.12.2003, p. 325-330.

Research output: Contribution to journalArticle

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