SS2.03 Phytoplankton Ecology Using Molecular Approach
Date: Thursday, June 13, 2002
Time: 3:00:00 PM
Location: Colwood
 
LitakerRW, National Ocean Service, NOAA, 101 Pivers Island Rd. , Beaufort, NC 28516-9722, , wayne_litaker@med.unc.edu
Vandersea, M, W, National Ocean Service, NOAA, 101 Pivers Island Rd. , Beaufort, NC 28516-9722, USA, mark.w.vandersea@noaa.gov
Kibler, S, R, National Ocean Service, NOAA, 101 Pivers Island Rd. , Beaufort, NC 28516-9722, , steve.kibler@noaa.gov
Tester, P, A, National Ocean Service, NOAA, 101 Pivers Island Rd. , Beaufort, NC 28516-9722, , pat.tester@noaa.gov
 
DETERMINING THE LIFE CYCLE STAGES OF HETEROTROPHIC DINOFLAGELLATES USING PEPTIDE NUCLEIC ACID PROBES
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Determining the life cycle of heterotrophic dinoflagellates can be difficult depending on the prey organism required as a food source. In certain instances, these prey organisms have life cycle stages that can be confused with those of the dinoflagellates. This presentation addresses how to avoid common pitfalls when determining heterotrophic dinoflagellate life cycles by using a systematic combination of nuclear staining, high resolution video microscopy, electron microscopy and in situ hybridization with fluorescently labeled peptide nucleic acid (PNA) probes. The use of a species-specific fluorescently labeled probe is particularly important when confirming whether the observed life cycle stages belong to the same species or not. PNA probes are well suited for determining life cycle stages in that they penetrate cell walls more easily and reach equilibrium more quickly than do labeled oligonucleotide probes. The presentation will also address the positive and negative controls that have to be included in the in situ hybridization experiments to correctly interpret the results.