RNA visualization in bacteria by fluorescence in situ hybridization

Jay H. Russell, Kenneth Charles Keiler

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Scopus citations

Abstract

Detecting localized RNA in bacteria is difficult due to the properties of RNA and the small size of the cell. Fluorescence in situ hybridization (FISH) has been an invaluable method for detecting and imaging RNA. In FISH, RNA is fixed in its native subcellular position through chemical cross-linking. An oligonucleotide probe conjugated to a fluorophore is annealed to the target RNA, and the target RNA/probe hybrid is visualized using fluorescence microscopy. This chapter describes the use of FISH to visualize tmRNA, a regulatory RNA required for trans-translation. The method can be adapted to visualize the localization of other regulatory and messenger RNAs as well.

Original languageEnglish (US)
Title of host publicationBacterial Regulatory RNA
Subtitle of host publicationMethods and Protocols
EditorsKenneth C. Keiler
Pages87-95
Number of pages9
DOIs
StatePublished - Jul 27 2012

Publication series

NameMethods in Molecular Biology
Volume905
ISSN (Print)1064-3745

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

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    Russell, J. H., & Keiler, K. C. (2012). RNA visualization in bacteria by fluorescence in situ hybridization. In K. C. Keiler (Ed.), Bacterial Regulatory RNA: Methods and Protocols (pp. 87-95). (Methods in Molecular Biology; Vol. 905). https://doi.org/10.1007/978-1-61779-949-5_7