Details zur Publikation

Kategorie Textpublikation
Referenztyp Zeitschriften
DOI 10.1101/pdb.prot5366
Titel (primär) Fluorescence in situ hybridization for the identification of environmental microbes
Autor Wendeberg, A.
Quelle Cold Spring Harbor Protocols
Erscheinungsjahr 2010
Department UMB
Band/Volume 2010
Heft 1
Seite von prot5366
Sprache englisch
Abstract

This protocol presents methods for the phylogenetic identification of microorganisms in environmental samples (e.g., water and sediments) by means of fluorescence in situ hybridization (FISH) with rRNA-targeted oligonucleotide probes, followed by signal amplification with catalyzed reporter deposition (CARD). In this procedure, FISH probes are conjugated with the enzyme horseradish peroxidase (HRP). After hybridization, the subsequent deposition of fluorescently labeled tyramides results in substantially higher signal intensities on target cells than after FISH with probes labeled directly with fluorochromes. This protocol describes the custom labeling of tyramides with different fluorochromes, as well as sample preparation and cell permeabilization strategies for various microbial cell wall types. A protocol for sequential multicolor CARD-FISH for the simultaneous detection of different phylogenetic groups is also presented.

dauerhafte UFZ-Verlinkung https://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=12394
Wendeberg, A. (2010):
Fluorescence in situ hybridization for the identification of environmental microbes
Cold Spring Harbor Protocols 2010 (1), prot5366 10.1101/pdb.prot5366