Application of high-throughput culturing method to a single seawater sample

A massive high-throughput dilution-to-extinction culturing method (HTC) was applied to a single surface seawater sample to isolate diverse marine heterotrophic bacteria. SSU rRNA gene sequences of isolated bacterial strains were phylogenetically analyzed and also compared to the result of amplicon pyrosequencing performed using DNA extracted from the seawater sample. Amplicon pyrosequencing was also applied to the pooled HTC culture broths to analyze the phylogenetic diversity of bacteria that grew as a minor fraction during HTC performed in this study.