Preparation of DNA suitable for PCR amplification from fresh or fixed single dinoflagellate cells

A method is described to prepare total DNA from single cells of dinoflagell ates, which can be used for PCR amplification. As model organisms, we used a stock strain of Alexandrium catenella and cells of Dinophysis acuminata h arvested from the Atlantic Ocean. Fresh grown cells or cells maintained in different preservatives were tested as sources for DNA preparation. The met hod used to prepare DNA combines physicochemical and enzymatic procedures o n cells embedded in agarose plugs or beans. The agarose pieces containing t he DNA were used to perform PCR amplification of a fragment of DNA containi ng a 5.8S rRNA, gene and the flanking internal transcribed spacers (ITS1 an d ITS2).