Locating RNAs in situ with FISH-STIC probes

John R. Sinnamon, Kevin Czaplinski

Research output: Chapter in Book/Report/Conference proceedingChapter

2 Scopus citations

Abstract

The location of a molecule within the cell often provides important clues to its function and regulation, therefore techniques to locate RNA within cells are vital tools to study noncoding RNA function. Fluorescence in situ hybridization (FISH) is a simple and reliable approach to locate RNAs in any cell type. Intracellular localization of RNA using FISH (RNA-FISH) requires resolution at the single cell and single molecule level which can be achieved using fluorescent-labeled nucleic acid antisense probes. S equential T agged and I ntertwined oligodeoxyribonucleotide C omplex (FISH-STIC) probes are a straightforward means for laboratories to design their own FISH probes that can be synthesized commercially. Here we provide a detailed protocol for applying FISH-STIC probes for in situ hybridization on cultured cells as a convenient and flexible method for localizing individual RNAs with many fluorophores using fluorescence microscopy.

Original languageEnglish (US)
Title of host publicationRegulatory Non-Coding RNAs
Subtitle of host publicationMethods and Protocols
PublisherSpringer New York
Pages137-148
Number of pages12
ISBN (Electronic)9781493913695
ISBN (Print)9781493913688
DOIs
StatePublished - Sep 20 2014
Externally publishedYes

Keywords

  • Fluorescence in situ hybridization
  • Oligodeoxyribonucleotide probe
  • RNA detection
  • RNA localization
  • RNA-FISH

ASJC Scopus subject areas

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)

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