RAPID ISOLATION OF HIGH-QUALITY RNA FROM SYMBIOTIC DINOFLAGELLATES

This report details a reliable and efficient RNA extraction protocol f or the symbiotic dinoflagellate Symbiodinium microadriaticum Freudenth al (Gymnodiniales, Dinophyceae). The method typically gives yields of 500 mu g total RNA from 0.4 g wet weight of algae, and, in comparison to current protocols, it is technically simple and less time consuming . This method isolates high-quality, intact RNA from in vine cultured as well as host-isolated cells, as demonstrated by spectrophotometry, gel electrophoresis, and northern analysis. The total RNA obtained was suitable for reverse transcription and PCR amplification of Symbiodin ium cDNAs. We have successfully applied our method to isolate total RN A from a different dinoflagellate, Amphidinium carterae Hulburt (Gymno diniales, Dinophyceae), found in symbiotic association with marine inv ertebrates.