title: Fluorescence in situ hybridization of small non-coding RNAs.
abstract: RNA FISH is a powerful method to detect specific RNAs in fixed cells. It allows both localization and quantification of RNA molecules within individual cells and tissues. Refined RNA FISH methods have also been developed to determine RNA transcription and degradation rates. This chapter describes an RNA FISH protocol that we developed in order to study the expression and localization of satellite III RNAs. This specific class of non-coding RNAs is expressed in response to various cellular stresses including heat shock. This protocol is based on the use of a biotinylated LNA probe subsequently detected by a streptavidin-Alexa Fluor(®) 488 conjugate. A protocol allowing efficient coupling of RNA FISH and protein detection by immunofluorescence is also described in this chapter.
authors: CNRS, UMR 7365 IMoPA, Université de Lorraine, 9 avenue de la Forêt de Haye, Vandoeuvre-lès-Nancy, 54506, France.
authors: Valentin
authors: V
authors: Vautrot
authors: Christelle
authors: C
authors: Aigueperse
authors: Christiane
authors: C
authors: Branlant
authors: Isabelle
authors: I
authors: Behm-Ansmant
mesh_terms: D005455:Fluorescent Antibody Technique; D006834:Hydrazines; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D015394:Molecular Structure; D011506:Proteins / Q000032:analysis*; D019244:RNA, Satellite / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D058727:RNA, Small Untranslated / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D019809:Streptavidin
chemical_list: C417561:Alexa 488 hydrazide; D006834:Hydrazines; D011506:Proteins; D019244:RNA, Satellite; D058727:RNA, Small Untranslated; D019809:Streptavidin