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Optimization of RNA Purification Method from Ecklonia cava Kjellman (Laminariales, Phaeophyceae)
Jong-Sung Ahn, Seon-Ock Woo, Jeong-Ha Kim, Yoon-Sik Oh, Jung-Hyun Oak, Seung-Shic Yum
Department of Biological Science, Sungkyunkwan University, Suwon 440-746 and 1Division of Life Science, Gyeongsang National University, Jinju 660-701, Korea
A more rapid and efficient method to extract RNA from Ecklonia cava Kjellman (Laminariales, Phaeophyceae) was introduced in this study. Each step of the procedure was evaluated and the optimal concentration of each chemical in the lysis solution was determined. Tissue pulverization with PVPP and β-mercaptoethanol in the lysis solution were not essential for RNA extraction of this species. The highest yield and purity of E. cava RNA were obtained by the lysis solution containing 1% CTAB, 1 M NaCl, 0.7% PVP, 10mM EDTA and 100mM Tris-Cl (pH 9.0). Approximately 8μg of RNA was obtained from 200 mg of ground tissue. The ratios of the absorbance at 260 nm and 280 nm were from 1.6 to 1.8 and those of at 230 nm and 260 nm were from 1.8 to 2.0. The extracted RNAs obtained in this study turned out to have a sufficient quality for cDNA synthesis.
CTAB, Ecklonia cava, EDTA, NaCl, PVP, RNA purification