An effective DNA extraction protocol for brown algae

Abstract

SUMMARY Successful extraction of total DNA from brown algae,which are generally polysaccharide and polyphenol rich,is often problematic using current methods. Persistentpolysaccharide and polyphenolic compounds canhinder further application of modern molecular tech-niques requisite to molecular-based evolutionary studies.Our broad and long-term research goals with fucaleantaxa, especially Sargassum , and problems with existingDNA extraction methods were an impetus to develop areliable DNA extraction method. Initial research estab-lished hexadecyltrimethylammonium bromide (CTAB)based total-DNA methods as the most viable for furtherempirical development. Several constituents effectiveat either complexing secondary compounds or creatinga reductive extraction environment were increased inconcentration or added to the extraction buffer. Theseseemingly minor changes resulted in the creation of ahighly reductive extraction buffer and effective total-DNA harvesting technique. We detail these modifica-tions and demonstrate the reliability of the modifiedprotocol with a variety of brown algae and tissue preser-vation methods. Such DNA is shown to be suitable fora variety of molecular techniques.Key words: brown algae, DNA extraction, CTABmethods, fucales, polyphenolic compounds,