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 journalArticle

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

Fingerprint

Bacterial DNA
Microarrays
DNA
Oligonucleotide Probes
Oligonucleotides
Ribosomal DNA
Oscillatoria
Nostoc
Microcystis
Anabaena
Synechococcus
Digoxigenin
Magnetic separation
Streptavidin
Biotin
Robots

Keywords

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

ASJC Scopus subject areas

  • Biotechnology
  • Microbiology

Cite this

Fluorescent detection of cyanobacterial DNA using bacterial magnetic particles on a MAG-microarray. / Matsunaga, Tadashi; Nakayama, Hideki; Okochi, Mina; Takeyama, Haruko.

In: Biotechnology and Bioengineering, Vol. 73, No. 5, 05.06.2001, p. 400-405.

Research output: Contribution to journalArticle

@article{722ac02c40fe4227aa6ad217a62b9ff8,
title = "Fluorescent detection of cyanobacterial DNA using bacterial magnetic particles on a MAG-microarray",
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.",
keywords = "Bacterial magnetic particle, MAG-microarray, Magnetic-separation robot, Oligonucleotide probe",
author = "Tadashi Matsunaga and Hideki Nakayama and Mina Okochi and Haruko Takeyama",
year = "2001",
month = "6",
day = "5",
doi = "10.1002/bit.1073",
language = "English",
volume = "73",
pages = "400--405",
journal = "Biotechnology and Bioengineering",
issn = "0006-3592",
publisher = "Wiley-VCH Verlag",
number = "5",

}

TY - JOUR

T1 - Fluorescent detection of cyanobacterial DNA using bacterial magnetic particles on a MAG-microarray

AU - Matsunaga, Tadashi

AU - Nakayama, Hideki

AU - Okochi, Mina

AU - Takeyama, Haruko

PY - 2001/6/5

Y1 - 2001/6/5

N2 - 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.

AB - 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.

KW - Bacterial magnetic particle

KW - MAG-microarray

KW - Magnetic-separation robot

KW - Oligonucleotide probe

UR - http://www.scopus.com/inward/record.url?scp=0035811090&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035811090&partnerID=8YFLogxK

U2 - 10.1002/bit.1073

DO - 10.1002/bit.1073

M3 - Article

C2 - 11320510

AN - SCOPUS:0035811090

VL - 73

SP - 400

EP - 405

JO - Biotechnology and Bioengineering

JF - Biotechnology and Bioengineering

SN - 0006-3592

IS - 5

ER -