Group‐specific primer and probe sets to detect methanogenic communities using quantitative real‐time polymerase chain reaction

Youngseob Yu; Changsoo Lee; Jaai Kim; Seokhwan Hwang

doi:10.1002/bit.20347

journal article Feb 04, 2005

Group‐specific primer and probe sets to detect methanogenic communities using quantitative real‐time polymerase chain reaction

Youngseob Yu Changsoo Lee Jaai Kim Seokhwan Hwang

Biotechnology and Bioengineering Vol. 89 No. 6 pp. 670-679 · Wiley

View at Publisher Save 10.1002/bit.20347

Abstract

Abstract

Real‐time polymerase chain reaction (PCR) is a highly sensitive method that can be used for the detection and quantification of microbial populations without cultivating them in anaerobic processes and environmental samples. This work was conducted to design primer and probe sets for the detection of methanogens using a real‐time PCR with the TaqMan system. Six group‐specific methanogenic primer and probe sets were designed. These sets separately detect four orders (
Methanococcales
,
Methanobacteriales
,
Methanomicrobiales
, and
Methanosarcinales
) along with two families (
Methanosarcinaceae
and
Methanosaetaceae
) of the order
Methanosarcinales
. We also designed the universal primer and probe sets that specifically detect the 16S rDNA of prokaryotes and of the domain
Bacteria
and
Archaea
, and which are fully compatible with the TaqMan real‐time PCR system. Target‐group specificity of each primer and probe set was empirically verified by testing DNA isolated from 28 archaeal cultures and by analyzing potential false results. In general, each primer and probe set was very specific to the target group. The primer and probe sets designed in this study can be used to detect and quantify the order‐level (family‐level in the case of
Methanosarcinales
) methanogenic groups in anaerobic biological processes and various environments. © 2005 Wiley Periodicals, Inc.

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Details

Published: Feb 04, 2005
Vol/Issue: 89(6)
Pages: 670-679
License: View

Authors

Cite This Article

Youngseob Yu, Changsoo Lee, Jaai Kim, et al. (2005). Group‐specific primer and probe sets to detect methanogenic communities using quantitative real‐time polymerase chain reaction. Biotechnology and Bioengineering, 89(6), 670-679. https://doi.org/10.1002/bit.20347

Group‐specific primer and probe sets to detect methanogenic communities using quantitative real‐time polymerase chain reaction

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