TECHNICAL NOTE Single-cell sequencing of dinoflagellate (Dinophyceae) nuclear ribosomal genes

Genetic sequences from dinoflagellates offer valuable information regarding taxonomies, phylogenies and population genetics that generally require the growth of these organisms in culture. We have developed a quick and simple method to obtain small and large subunit ribosomal gene sequences from dinoflagellates using single cells. This method, based on freeze–thaw cell lysis and a simple two-step polymerase chain reaction, provides template for sequencing in 6–8 h. We have sequenced five dinoflagellate species, including unculturable Dinophysis and Ceratium species, using fresh and frozen samples.