寡核苷酸探针的荧光原位杂交双重标记（DOPE-FISH）提高了信号强度并增加了 rRNA 的可及性。
Double labeling of oligonucleotide probes for fluorescence in situ hybridization (DOPE-FISH) improves signal intensity and increases rRNA accessibility.
机构信息
Department of Microbial Ecology, University of Vienna, Vienna, Austria.
出版信息
Appl Environ Microbiol. 2010 Feb;76(3):922-6. doi: 10.1128/AEM.02456-09. Epub 2009 Dec 4.
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.
摘要
荧光原位杂交（FISH）技术使用单标记的 rRNA 靶向寡核苷酸探针，广泛应用于环境或临床标本中微生物的直接鉴定。本文显示，用 5' 、 3' 双标记探针替代单标记寡核苷酸探针，至少可以将 FISH 信号强度提高一倍，而不会产生特异性问题。此外，Cy3 双标记探针显著增加了 rRNA 靶位的原位可及性，从而为探针设计提供了更大的灵活性。
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