Demonstration of extraction and PCR amplification of DNA from phytoplankton of lakes with high humic acid content

Abstract

Lakes with high levels of dissolved organic matter are common in northern temperate regions. These habitats are sensitive to environmental and climate change, but the molecular ecology and eco-physiology of their phytoplankton have been under studied, in part because the DOM interferes with molecular and biochemical analyses of these populations. We developed a procedure which uses polyvinylpyrrolidone to adsorb interfering material during the extraction of nucleic acids from these populations. This procedure does not depend on enzymatic lysis and yields DNA of sufficient quality to serve as a template for PCR amplification of specific genetic sequences. In particular, we detected fragments of the conserved rbcL gene encoding the large subunit of the Rubisco enzyme from the phytoplankton of lakes with high DOM. Our extraction procedure may be useful to other workers studying similar populations.