DNA EXTRACTION CONDITIONS FROM PORPHYRA-PERFORATA USING LICL

A rapid and economical method of DNA extraction from a red seaweed Porphyra perforata J. Agardh has been developed by the use of lithium chloride. This paper describes the optimization of extraction conditions. Heat treatment of tissues in a solution (0.8 M LiCl, 0.6% Sarkosyl, 10 mM EDTA, 0.2% PVPP, 5% beta-mercaptoethanol, pH 9.0) at 55 degrees C for 10 min extracts DNA that is of sufficient quality to be used as a template for PCR amplification. Total DNA yield was approximately 30 to 50 mu g g(-1) of partially dried tissue. Total RNA yield was approximately 400 mu g g(-1) of partially dried tissue. Carbohydrate was contained as approximately 40 to 90 mM (expressed as glucose equivalents) from 1 g tissue, and protein contamination calculated as the O.D. 260/280 ratio was in the range of 1.4 to 1.7. The DNA was characterized by high molecular weight larger than 50 kb.