A protocol for PCR in situ hybridization of hyphomycetes

doi:10.2436/im.v1i3.43

International Microbiology, Vol 1, No 3 (1998)

A protocol for PCR in situ hybridization of hyphomycetes

Katja Sterflinger, Wolfgang E. Krumbein, Andreas Schwiertz

Abstract

A protocol for application of Polymerase Chain Reaction (PCR) in situ hybridization for the detection of hyphomycetes is presented. The experiments are exemplary carried out with strains of the genera Penicillium and Cladosporium. The small ribosomal subunit is amplified in situ by PCR using fungal specific primers. The amplicon is used as target region for a fluorescein-marked probe. The permeability of the fungal cell wall for the primers and the probe can be successfully achieved by enzymatic treatment with ß-glucanase. The protocol can be used as a basis for further development of in situ hybridization with taxon specific probes

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