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JOURNAL OF PLANKTON RESEARCH | VOLUME 5 | NUMBER 2 | PAGES 253-261 | 1983
© Oxford University Press

research-article

A rapid technique for the determination of RNA and DNA in marine phytoplankton

Steven S. Thoresen, John R. Clayton, Jr.1, Quay Dortch2 and S.I. Ahmed

School of Oceanography, WB-10, University of Washington Seattle, WA 98195, USA

Received on June 1, 1982; accepted on November 1, 1983

An adaptation of the ethidium bromide technique for the analysis of nucleic acids is presented for marine phytoplankton. The method involves an initial homogenization of cells in phosphate buffered saline, followed by incubation of subsamples of the cell homogenate in the presence and absence of ribonudease. Quantities of DNA and DNA + RNA in the respective sub-samples are then determined by reaction with ethidium bromide. An evaluation is made of appropriate levels of bentonite required in the assay to inhibit endogenous cellular ribonucleases. Two nucleoprotein dissociating agents, pronase and heparin, are also investigated for their capacities to enhance nucleic add fluorescence yield. The final recommended method resulted in maximum measured levels of RNA and DNA in phytoplankton samples tested. The method can be rapidly performed, involves a minimum amount of sample manipulation, and yields numbers having a high degree of precision.

1Current address: School of Fisheries, University of Washington, WH-l0, Seattle, WA 98195, USA.

2Current address: Bigelow Laboratory for Ocean Sciences, West Boothbay Harbor, ME 04575, USA.


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[Abstract] [Full Text]


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