title: Tracking DNA and RNA sequences at high resolution.
abstract: In this chapter, the basic principles and protocols of the electron microscopical detections of specific DNA and RNA sequences are described. We focused primarily on a comparison of various methods of electron microscopy in situ hybridization (EM-ISH) with respect to their sensitivity and the structural preservation of the sample with the aim of helping the readers select the appropriate hybridization protocol. As the post-embedding EM-ISH most frequently represents the optimal choice, the protocol for the post-embedding EM-ISH approach is described in detail. Concurrently, the alternative methods based on the enzymatic synthesis of the labeled nucleic acids chains that can be used for the detection of DNA or RNA molecules in situ are mentioned. In this respect, the technique enabling the enzymatic detection of the polyadenylated RNA sequences is described in detail.
authors.affiliation: Institute of Cellular Biology and Pathology, 1st Faculty of Medicine, Charles University in Prague, Prague, Czech Republic.
authors.forename: Dušan
authors.initials: D
authors.lastname: Cmarko
authors.forename: Anna
authors.initials: A
authors.lastname: Ligasová
authors.forename: Karel
authors.initials: K
authors.lastname: Koberna
mesh_terms: D004247:DNA / Q000737:chemistry*; D017403:In Situ Hybridization / Q000379:methods*; D008854:Microscopy, Electron / Q000379:methods*; D008867:Microtomy / Q000379:methods; D012313:RNA / Q000737:chemistry*
chemical_list: D012313:RNA; D004247:DNA