Abstract
Fluorescence in situ hybridization (FISH) with singly labeled rRNA-targeted oligonucleotide probes is widely applied for direct identification of microbes in the environment or in clinical specimens. Here we show that a replacement of singly labeled oligonucleotide probes with 5'-, 3'-doubly labeled probes at least doubles FISH signal intensity without causing specificity problems. Furthermore, Cy3-doubly labeled probes strongly increase in situ accessibility of rRNA target sites and thus provide more flexibility for probe design.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Conserved Sequence / genetics
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Escherichia coli / genetics
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Evolution, Molecular
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Flow Cytometry
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Fluorescence
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Fluorescent Dyes / chemistry*
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In Situ Hybridization, Fluorescence / methods*
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Nucleic Acid Conformation
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Oligonucleotide Probes / chemistry*
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Phylogeny
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RNA, Bacterial / analysis*
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RNA, Ribosomal / analysis*
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RNA, Ribosomal, 16S / analysis
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RNA, Ribosomal, 18S / analysis
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Sequence Alignment
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Species Specificity
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Thermodynamics
Substances
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Fluorescent Dyes
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Oligonucleotide Probes
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RNA, Bacterial
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RNA, Ribosomal
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RNA, Ribosomal, 16S
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RNA, Ribosomal, 18S