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Published 2003

Read in Norwegian

Publication details

Journal : Preparative Biochemistry & Biotechnology , vol. 33 , p. 197–208 , 2003

International Standard Numbers :
Printed : 1082-6068
Electronic : 1532-2297

Publication type : Academic article

Contributors : Rudi, Knut; Treimo, Janneke Regine; Nissen, Hilde; Vegarud, Gerd Elisabeth

Issue : 3

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Kjetil Aune
Chief Librarian


Analyses of complex microbial communities are becoming increasingly important. Bottlenecks in these analyses, howeve, are the tools to actually describe the biodiversity. Novel protocols for DNA array based analyses of microbial communities are presented. In these protocols, the specificity obtained by sequence-specific labeling of DNA probes is combined with the possibility of detecting several different probes simultaneously by DNA array hybridization. The gene encoding 16S ribosomal RNA was chosen as the target in these analyses. This gene contains both universally conserved regions, and regions with relatively high variability. The universally conserced regions are used for PCR amplification primers, while the variable regions are used for specific proves. Arrays prepared on positively charges nylon membranes and coated glass slides were compared. The advantage of using membranes is that chromatogenic signal amplification can be used for the detection. Furthermore, the chromogenic detection does not require any sophisticated equipment. The advantage of the glass slides is that multiple fluorescence colors can be detected simultaneously, ant that internal controls can be used for normalization. This approach is also suited for high throughput screenings.