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-microm-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.

journal_name

Biotechnol Bioeng

authors

Matsunaga T,Nakayama H,Okochi M,Takeyama H

doi

10.1002/bit.1073

subject

Has Abstract

pub_date

2001-06-05 00:00:00

pages

400-5

issue

5

eissn

0006-3592

issn

1097-0290

pii

10.1002/bit.1073

journal_volume

73

pub_type

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