title: Colorimetric in situ hybridization (CISH) with digoxigenin-labeled oligonucleotide probes in autofluorescent hyphomycetes.
abstract: We used digoxigenin-labeled probes for in situ hybridization of hyphomycetes to replace the commonly used fluorescent proof of probe binding by a colorimetric reaction. The resulting blue-purple, intracellular precipitate could be easily detected by light microscopy, and thus presented a promising method to overcome autofluorescence of fungal material and substratum. Optimal cell fixation and permeabilization procedures, as well as hybridization conditions were developed on the example of two different hyphomycetes: Phialophora sp. and Cartapip, a colorless mutant of Ophiostoma piliferum (Agra Sol).
authors.affiliation: Department of Geomicrobiology, Institute of Chemistry and Biology of the Marine Environment, Carl von Ossietzky-University, Oldenburg, Germany. susanne.schroeder@mail.uni-oldenburg.de
authors.forename: S
authors.initials: S
authors.lastname: Schröder
authors.forename: M
authors.initials: M
authors.lastname: Hain
authors.forename: K
authors.initials: K
authors.lastname: Sterflinger
mesh_terms: D011232:Chemical Precipitation; D003124:Colorimetry / Q000379:methods*; D004271:DNA, Fungal / Q000032:analysis*; D004076:Digoxigenin; D005453:Fluorescence; D017403:In Situ Hybridization / Q000379:methods*; D003904:Mitosporic Fungi / Q000235:genetics / Q000302:isolation & purification*; D015345:Oligonucleotide Probes; D010676:Phialophora / Q000235:genetics / Q000302:isolation & purification; D016707:Tissue Fixation
chemical_list: D004271:DNA, Fungal; D015345:Oligonucleotide Probes; D004076:Digoxigenin