title: Fluorescence In Situ Hybridization in Oat.
abstract: This chapter describes methods to detect gene loci or gene transcripts by fluorescence labeling. Fluorescence in situ hybridization (FISH) can be used to identify the positions of genes or BACs or the distribution of repetitive sequences on metaphase chromosomes as well as the identification of alien chromosomes. It enables the identification of gene loci and active transcription sites in interphase nuclei and also the localization of cellular transcripts. The protocols here deal with the production of DNA and RNA probes, the preparation of oat metaphase spreads and root tissue sections, the subsequent hybridization, post-hybridization washes, and detection by immunofluorescence.
authors.affiliation: Cell and Developmental Biology, John Innes Centre, Colney Lane, Norwich, NR4 7UH, UK. eva.wegel@jic.ac.uk.
authors.forename: Eva
authors.initials: E
authors.lastname: Wegel
mesh_terms: D018554:Avena / Q000235:genetics*; D002874:Chromosome Mapping; D032461:Chromosomes, Plant; D015870:Gene Expression; D056426:Genetic Loci; D017404:In Situ Hybridization, Fluorescence / Q000379:methods; D008677:Metaphase / Q000235:genetics; D015341:Nucleic Acid Probes; D012333:RNA, Messenger / Q000235:genetics
chemical_list: D015341:Nucleic Acid Probes; D012333:RNA, Messenger
keywords: FISH; Immunofluorescence; In vitro transcription; Metaphase spreads; Nick translation; Wax embedding