title: RNA in situ hybridization in Arabidopsis.
abstract: RNA in situ hybridization using digoxigenin-labeled riboprobes on tissue sections is a powerful technique for revealing microscopic spatial gene expression. Here, we describe an in situ hybridization method commonly practiced in Arabidopsis research labs. The highly stringent hybridization condition eliminates the usage of Ribonlucease A and gives highly specific signals. This also allows the use of longer probes which enhance signal strength without cross hybridization to closely related genes. In addition, using spin columns in template and riboprobe purification greatly reduces background signals.
authors.affiliation: Department of Biology, University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: Miin-Feng
authors.initials: MF
authors.lastname: Wu
authors.forename: Doris
authors.initials: D
authors.lastname: Wagner
mesh_terms: D017360:Arabidopsis / Q000235:genetics*; D003890:Desiccation; D004076:Digoxigenin / Q000737:chemistry; D000431:Ethanol / Q000737:chemistry; D005404:Fixatives / Q000737:chemistry; D005557:Formaldehyde / Q000737:chemistry; D017403:In Situ Hybridization / Q000379:methods*; D007202:Indicators and Reagents / Q000737:chemistry; D012313:RNA / Q000235:genetics*; D015347:RNA Probes; D018749:RNA, Plant / Q000235:genetics*; D013194:Staining and Labeling; D016610:Tissue Embedding / Q000379:methods; D014158:Transcription, Genetic
chemical_list: D005404:Fixatives; D007202:Indicators and Reagents; D015347:RNA Probes; D018749:RNA, Plant; D005557:Formaldehyde; D000431:Ethanol; D012313:RNA; D004076:Digoxigenin