Evaluation of enzymatic cell treatments for application of CARD-FISH to methanogens.

Evaluation of enzymatic cell treatments for application of CARD-FISH to methanogens. Research Abstract Details 

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Evaluation of enzymatic cell treatments for application of CARD-FISH to methanogens. Abstract Text:

Kengo Kubota,Hiroyuki Imachi,Shuji Kawakami,Kohei Nakamura,Hideki Harada,Akiyoshi Ohashi,Kengo Kubota,Hiroyuki Imachi,Shuji Kawakami,Kohei Nakamura,Hideki Harada,Akiyoshi Ohashi,

Several enzymatic permeabilization protocols (utilizing lysozyme, proteinase K, achromopeptidase, or recombinant pseudomurein endopeptidase [PeiW]) were evaluated for application of in situ hybridization with horseradish peroxidase-labeled oligonucleotide probes and catalyzed reporter deposition (CARD-FISH) to methanogens. In this study, twelve methanogens were selected that have typical cell surface structures: pseudomurein, surface layer, methanochondroitin and sheath. Among the treatments tested, PeiW treatment was observed to be the most effective one, although methanogens having a sheath were stained heterogeneously and methanogens having methanochondroitin were not permeabilized. On the other hand, lysozyme, proteinase K, and achromopeptidase treatments were ineffective or caused cell-lysis, resulting in weak or no signals. Applicability of PeiW treatment was further evaluated using an anaerobic granular sludge sample. The detection rate of Archaea by CARD-FISH increased remarkably after the treatment. Based on the results obtained in this study, we propose PeiW treatment as a novel permeabilization method for CARD-FISH application to methanogens.

Evaluation of enzymatic cell treatments for application of CARD-FISH to methanogens. Publishing Authors By Initials

K Kubota,H Imachi,S Kawakami,K Nakamura,H Harada,A Ohashi,K Kubota,H Imachi,S Kawakami,K Nakamura,H Harada,A Ohashi,

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Evaluation of enzymatic cell treatments for application of CARD-FISH to methanogens. Journal Published:

PUBLICATION TYPE: Journal Article

Journal: Journal of microbiological methods

VOLUME: 72

Page Numbers: 54-9

Journal Abbreviation: J. Microbiol. Methods

ISSN: 0167-7012

DAY: 26

MONTH: 10

YEAR: 2007

Evaluation of enzymatic cell treatments for application of CARD-FISH to methanogens. Information

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LANGUAGE: eng

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Grant and Affiliation Information for Evaluation of enzymatic cell treatments for application of CARD-FISH to methanogens.

AFFILIATION: Department of Environmental Systems Engineering, Nagaoka University of Technology, Nagaoka, Japan; Department of Civil and Environmental Engineering, Tohoku University, Sendai, Japan.

Country: Netherlands

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MEDLINETA: J Microbiol Methods

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