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The Korean Society of Phycology (한국조류학회(藻類))
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Optimization of RNA Purification Method from Ecklonia cava Kjellman (Laminariales, Phaeophyceae)

  • Published : 2004.06.30
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Abstract

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.

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References

  1. Ahn M.-J.,Yoon K-D., Kim cv, Min S.-Y.,Kim Y.-V., Kim H.J., Kim J.H., Shin c.c, Lee C.-K, Kim T.G., Kim S.H., Huh H. and Kim J. 2002. Inhibition of HIV-l reverse transcriptase and HIV-l invertase and antiviral activity of Korean seaweed extracts. J. Appl. Phycol. 14: 325-329 https://doi.org/10.1023/A:1022192329471
  2. Ahn M.-J., Yoon K-D., Min S.-Y., Lee J.S., Kim J.H., Kim T.G., Kim S.H., Kim, N.-G. and Kim, J. 2004. Inhibition of HIV-l reverse transcriptase and protease by phlorotannins from a brown alga, Ecklonia cava. Biol. Pharm. Bull. 27: 544-547 https://doi.org/10.1248/bpb.27.544
  3. Apt K.E., Clendennen S.K, Powers D.A and Grossman A.R. 1995. The gene family encoding the fucoxanthin chlorophyll proteins from the brown alga Macrocystis pyrifera. Mol. Gen. Genet. 246: 455-464 https://doi.org/10.1007/BF00290449
  4. Hong Y.-K, Kim S.-D., Polne-Fuller M. and Gibor A. 1995a. DNA extraction conditions from Porphyra perforata using LiCI. J. Appl. Phycol. 7: 101-107 https://doi.org/10.1007/BF00693055
  5. Hong Y.-K, Sohn C.H. Polne-Fuller M. and Gibor A 1995b. Differential display of tissue-specific messenger RNAs in Porphyra perforata (Rhodophyta) thallus. J. Phycol. 31: 640-643 https://doi.org/10.1111/j.1529-8817.1995.tb02560.x
  6. Kitade Y., Yamazaki S. and Saga N. 1996. A method for extraction of high molecular weight DNA from the macroalga Porphyra yezoensis (Rhodophyta). J. Phycol. 32: 496-498 https://doi.org/10.1111/j.0022-3646.1996.00496.x
  7. Lee Y.K. and Lee H.K. 2003. A simple method for DNA extraction from red algae. Algae18: 65-69 https://doi.org/10.4490/ALGAE.2003.18.1.065
  8. Mayes c, Saunders G.W., Tan I.H. and Druehl L.D. 1992.DNA extraction methods for kelp (Laminariales) tissue. J. Phycol. 28: 712-716 https://doi.org/10.1111/j.0022-3646.1992.00712.x
  9. Sambrook J. and Russell D.W. 2001. Moclecular cloning: a laboratory manual. 3rd ed. Cold Spring Harbor Laboratory Press, New York
  10. Su X. and Gibor A 1988. A method for RNA isolation from marine macro-algae. Anal. Biochem. 174: 650-657 https://doi.org/10.1016/0003-2697(88)90068-1

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