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  The effect of nucleobase-specific fluorescence quenching on in situ hybridization with rRNA-targeted oligonucleotide probes

Behrens, S., Fuchs, B. M., & Amann, R. (2004). The effect of nucleobase-specific fluorescence quenching on in situ hybridization with rRNA-targeted oligonucleotide probes. Systematic and Applied Microbiology, 27(5), 565-572.

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Behrens4.pdf (Publisher version), 195KB
 
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 Creators:
Behrens, S.1, Author           
Fuchs, B. M.1, Author           
Amann, R.1, Author           
Affiliations:
1Department of Molecular Ecology, Max Planck Institute for Marine Microbiology, Max Planck Society, ou_2481696              

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Free keywords: fluorescence in situ hybridization (FISH) photoinduced electron transfer; oligonucleotide probes; nucleobase-specific quenching; fluorescent dyes; 6-carboxyfluorescein (6-FAM)
 Abstract: Oligonucleotide probes labeled with fluorescent dyes are used in a variety of in situ applications to detect specific DNA or RNA molecules. It has been described that probe fluorescence might be quenched upon hybridization in a sequence specific way. Here, a set of 17 oligonuleotides labeled with 6-carboxyfluorescein was used to examine the relevance of nucleotide specific quenching for fluorescence in situ hybridization (FISH) to whole fixed bacterial cells. Probes quenched upon hybridization to a guanine-rich region of purified RNA in solution were not quenched upon FISH. Among other factors the high protein concentration within cells may prevent quenching of probe fluorescence in situ.

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Language(s): eng - English
 Dates: 2004-12-01
 Publication Status: Issued
 Pages: 8
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: eDoc: 222375
ISI: 000224248400008
 Degree: -

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Title: Systematic and Applied Microbiology
  Other : System. Appl. Microbiol.
Source Genre: Journal
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Publ. Info: Stuttgart : Urban & Fischer
Pages: - Volume / Issue: 27 (5) Sequence Number: - Start / End Page: 565 - 572 Identifier: ISSN: 0723-2020
CoNE: https://pure.mpg.de/cone/journals/resource/954928582871