| Detection of prokaryotic cells with fluorescence in situ hybridization. | |
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MedLine Citation:
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PMID: 20809326 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Fluorescence in situ hybridization with rRNA targeted oligonucleotide probes is nowadays one of the core techniques in microbial ecology, allowing the identification and quantification of microbial cells in environmental samples in situ. Next to the classic FISH protocol, which uses fluorescently monolabelled probes, the more sensitive CARD-FISH (also known as TSA-FISH), which involves an enzyme catalyzed signal amplification step, is becoming increasingly popular. This chapter describes protocols for both methods. While classic FISH has the advantage of being relatively cheap and easy to do on morphologically diverse samples, CARD-FISH offers a significantly higher sensitivity, allowing the detection of slow growing or metabolically inactive cells, which are below the detection limit of classic FISH. The drawback here is the considerably higher price for the probes and advanced cell fixation and permeabilization requirements that have to be optimized for different target cells. |
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Authors:
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Katrin Zwirglmaier |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Methods in molecular biology (Clifton, N.J.) Volume: 659 ISSN: 1940-6029 ISO Abbreviation: Methods Mol. Biol. Publication Date: 2010 |
Date Detail:
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Created Date: 2010-09-02 Completed Date: 2010-12-14 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9214969 Medline TA: Methods Mol Biol Country: United States |
Other Details:
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Languages: eng Pagination: 349-62 Citation Subset: IM |
Affiliation:
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British Antarctic Survey, Cambridge, UK. k.zwirglmaier@warwick.ac.uk |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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In Situ Hybridization, Fluorescence
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methods* Oligonucleotide Probes / genetics, metabolism Prokaryotic Cells / cytology*, metabolism |
| Chemical | |
Reg. No./Substance:
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0/Oligonucleotide Probes |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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