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Applied and Environmental Microbiology, November 2000, p. 5066-5072, Vol. 66, No. 11
Deep-sea Microorganisms Research Group, Japan
Marine Science & Technology Center, Yokosuka 237-0061, Japan
Received 1 June 2000/Accepted 14 August 2000
We describe a rapid, reproducible, and sensitive method for
detection and quantification of archaea in naturally occurring microbial communities. A domain-specific PCR primer set and a domain-specific fluorogenic probe having strong and weak selectivity, respectively, for archaeal rRNA genes (rDNAs) were designed. A universal PCR primer set and a universal fluorogenic probe for both
bacterial and archaeal rDNAs were also designed. Using these primers
and probes, we demonstrated that detection and quantification of
archaeal rDNAs in controlled microbial rDNA assemblages can be
successfully achieved. The system which we designed was also able to
detect and quantify archaeal rDNAs in DNA samples obtained not only
from environments in which thermophilic archaea are abundant but also
from environments in which methanogenic archaea are abundant. Our
findings indicate that this method is applicable to culture-independent molecular analysis of microbial communities in various environments.
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Rapid Detection and Quantification of Members of the Archaeal
Community by Quantitative PCR Using Fluorogenic Probes
*
Corresponding author. Mailing address: Deep-sea
Microorganisms Research Group, Japan Marine Science & Technology
Center, 2-15 Natsushima-cho, Yokosuka 237-0061, Japan. Phone:
81-468-67-3894. Fax: 81-468-66-6364. E-mail:
kent{at}jamstec.go.jp.
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