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- Title
IDENTIFICATION OF THE CLASS PRYMNESIOPHYCEAE AND THE GENUS <em>PHAEOCYSTIS</em> WITH RIBOSOMAL RNA-TARGETED NUCLEIC ACID PROBES DETECTED BY FLOW CYTOMETRY.
- Authors
Lange, Martin; Guillou, Laure; Vaulot, Daniel; Simon, Nathalie; Amann, Rudolf I.; Ludwig, Wolfgang; Medlin, Linda K.
- Abstract
Target regions specific for the class Prymnesiophyceae and the genus Phaeocystis (Har.) Lag., were identified from 18S ribosomal RNA coding regions, and two complementary probes were designed (PRYMN01 and PHAEO01). Detection of whole cells hybridized with these probes labeled with fluorescein isothiocyanate was difficult using epifluorescence microscopy because autofluorescence of the chlorophylls seriously interfered with the fluorescence of the probes. In contrast, flow cytometry proved very useful to detect and quantify the fluorescence of the hybridized cells. Hybridization conditions were opotimized, especially with respect to formamide concentration. Both probes were tested on a large array of both target and nontarget strains. Positive and negative controls were also analyzed. Specificity was tested by adding a competing nonlabled probe. Whereas probe PHAEO01 appeared less specific and must be used with stringent positive and negative controls.
- Subjects
PRYMNESIOPHYCEAE; NUCLEIC acid probes; ALGAE; RIBOSOMES; RNA; FLOW cytometry
- Publication
Journal of Phycology, 1996, Vol 32, Issue 5, p858
- ISSN
0022-3646
- Publication type
Academic Journal
- DOI
10.1111/j.0022-3646.1996.00858.x