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An effective DNA extraction protocol for brown algae
Authors:Naomi Phillips †  Celia M Smith  Clifford W Morden
Institution:Department of Botany, University of Hawaii, Honolulu, HI 96822, USA;Center for Conservation Research and Training, University of Hawaii, Honolulu, HI 96822, USA.
Abstract:Successful extraction of total DNA from brown algae, which are generally polysaccharide and polyphenol rich, is often problematic using current methods. Persistent polysaccharide and polyphenolic compounds can hinder further application of modern molecular techniques requisite to molecular‐based evolutionary studies. Our broad and long‐term research goals with fucalean taxa, especially Sargassum, and problems with existing DNA extraction methods were an impetus to develop a reliable DNA extraction method. Initial research established hexadecyltrimethylammonium bromide (CTAB) based total‐DNA methods as the most viable for further empirical development. Several constituents effective at either complexing secondary compounds or creating a reductive extraction environment were increased in concentration or added to the extraction buffer. These seemingly minor changes resulted in the creation of a highly reductive extraction buffer and effective total‐ DNA harvesting technique. We detail these modifications and demonstrate the reliability of the modified protocol with a variety of brown algae and tissue preservation methods. Such DNA is shown to be suitable for a variety of molecular techniques.
Keywords:brown algae  DNA extraction  CTAB methods  fucales  polyphenolic compounds              Sargassum
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