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A simple and innovative method for species identification of phytoplankton cells on minute quantities of DNA
Article first published online: 28 JUN 2010
DOI: 10.1111/j.1758-2229.2010.00164.x
© 2010 Society for Applied Microbiology and Blackwell Publishing Ltd
Additional Information
How to Cite
Masseret, E., Enquebecq, M., Laabir, M., Genovesi, B., Vaquer, A. and Avarre, J.-C. (2010), A simple and innovative method for species identification of phytoplankton cells on minute quantities of DNA. Environmental Microbiology Reports, 2: 715–719. doi: 10.1111/j.1758-2229.2010.00164.x
Publication History
- Issue published online: 21 NOV 2010
- Article first published online: 28 JUN 2010
- Received 4 September, 2009; accepted 22 February, 2010.
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Summary
Dinoflagellates belonging to the genus Alexandrium are often involved in harmful algal blooms. Their ecological exploration is thus essential to increase our knowledge on these toxic events. Yet, population genetic studies, taxonomic identification and environmental monitoring are hampered by major constraints: the necessity to establish monoclonal cultures from environmental samples and the sensitivity of available molecular tools. The present work describes a very simple and sensitive method for extraction and amplification of DNA at the infra-single-cell level. Its on-slide format allows for easy visual control of both quality and quantity of the templates. Combined with a semi-multiplex PCR protocol designed on the 18S-28S rDNA-ITS region of Alexandrium catenella and Alexandrium tamarense, this procedure allowed the identification and discrimination of these species from both monoclonal cultures and natural samples.