Fluorescent detection of cyanobacterial DNA using bacterial magnetic particles on a MAG-microarray

Tadashi Matsunaga, Hideki Nakayama, Mina Okochi, Haruko Takeyama

Research output: Contribution to journalArticlepeer-review

41 Citations (Scopus)

Abstract

Bacterial magnetic particles (BMPs) were used for the identification of cyanobacterial DNA. Genus-specific oligonucleotide probes for the detection of Anabaena spp., Microcystis spp., Nostoc spp., Oscillatoria spp., and Synechococcus spp. were designed from the variable region of the cyanobacterial 16S rDNA of 148 strains. These oligonucleotide probes were immobilized on BMPs via streptavidin-biotin conjugation and employed for magnetic-capture hybridization against digoxigenin-labeled cyanobacterial 16S rDNA. Bacterial magnetic particles were magnetically concentrated, spotted in 100-μm-size microwell on MAG-microarray, and the fluorescent detection was performed. This work details the development of an automated technique for the magnetic isolation, the concentration of hybridized DNA, and the detection of specific target DNA on MAG-microarray. The entire process of hybridization and detection was automatically performed using a magnetic-separation robot and all five cyanobacterial genera were successfully discriminated.

Original languageEnglish
Pages (from-to)400-405
Number of pages6
JournalBiotechnology and bioengineering
Volume73
Issue number5
DOIs
Publication statusPublished - 2001 Jun 5
Externally publishedYes

Keywords

  • Bacterial magnetic particle
  • MAG-microarray
  • Magnetic-separation robot
  • Oligonucleotide probe

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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