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The Korean Society of Phycology (한국조류학회(藻類))
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A Simple Method for Extraction of High Molecular Weight DNA fromPorphyra Tenera (Rhodophyta) Using Diatomaceous Earth

  • Kim, Tae-Hoon (Department of Molecular Biology, Busan National University) ;
  • Hwang, Mi-Sook (Seaweed Research Center, South Sea Fisheries Research Institute, National Fisheries Research and Development Institute) ;
  • Song, Ju-Dong (Department of Molecular Biology, Busan National University) ;
  • Oh, Min-Hyuk (Department of Molecular Biology, Busan National University) ;
  • Moon, Yong-Hwan (Department of Molecular Biology, Busan National University) ;
  • Chung, Ik-Kyo (Department of Marine Science, Busan National University) ;
  • Rhew, Tae-Hyoung (Department of Biology, Busan National University) ;
  • Lee, Choon-Hwan (Department of Molecular Biology, Busan National University)
  • Published : 2006.06.30
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Abstract

The innate soluble polysaccharides and phenolic compounds of marine macroalgae are serious contaminants which interfere with experimental procedures such as restriction enzyme digestion, polymerase chain reaction (PCR) and other enzymatic reactions using extracted DNA samples. The viscous polysaccharides are co-precipitated with DNA samples by isopropanol or ethanol precipitation in conventional experiment. To overcome the problem, a method for the isolation of high molecular weight DNA from Porphyra tenera is developed with the application of diatomaceous earth column. The isolated DNAs by this method were about 50-100 kb in size and could be digested well with restriction enzymes. The nuclease activity seemed to be minimal, and high reproducibility in the arbitrary primed PCR for RAPD analyses was a distinctive feature. These results suggest that this method is very efficient in isolating nucleic acid from macroalgae including Porphyra.

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References

  1. Alberto F., Santos R. and Leitão J.M. 1997. DNA extraction and RAPD markers to access the genetic similarity among Gelidium sesquipedale (Rhodophyta) populations. J. Phycol. 33: 706-710 https://doi.org/10.1111/j.0022-3646.1997.00706.x
  2. Araki S., Sakurai T., Oohusa T. and Sato N. 1992. Comparative restriction endonuclease analysis of rhodoplast DNA from different species of Porphyra (Bangiales, Rhodophyta). Nippon Suisan Gakkaishi 58: 477-480 https://doi.org/10.2331/suisan.58.477
  3. Atyeo R.F., Oxberry S.L., Combs B.G. and Hampson D.J. 1998. Development and evaluation of polymerase chain reaction tests as an aid to diagnosis of swine dysentery and intesti-nal spirochaetosis. Lett. Appl. Microbiol. 26: 126-130 https://doi.org/10.1046/j.1472-765X.1998.00294.x
  4. Bold H.C. and Wynne M.J. 1978. Introduction to the algae. Structure and reproduction. Prentice-Hall. Englewood Cliffs. NJ
  5. Demeke T. and Adams R.P. 1992. The effects of plant polysaccharides and buffer additives on PCR. Bio Techniques 12:332-334
  6. Do N. and Adams R.P. 1991. A simple technique for removing plant polysaccharide contaminants from DNA. Bio Techniques 10: 162-166
  7. Hong Y.-K., Sohn C.H., Lee K.W. and Kim H.G. 1997. Nucleic acid extraction from seaweed tissues for polymerase chain reaction. J. Mar. Biotechnol. 5: 95-99
  8. Kamenetzky L., Canova S.G., Guarnera E.A. and Rosenzvit M.C. 2000. Echinococcus granulosus: DNA extraction from germinal layers allows strain determination in fertile and nonfertile hydatid cysts. Exp. Parasitol. 95: 122-127 https://doi.org/10.1006/expr.2000.4518
  9. Kim K.-S. and Pallaghy C.K. 1996. Purification of plasmid DNA (miniprep) with high yields using diatomaceous earth. U.S.DeptCommerce/NOAA/NMFS/NWFSC/Molecular biology protocol (http://www.nwfsc.noaa.gov/protocols/dnaprep.html)
  10. Kim S.H., Lee Y.K., Hong C.B. and Lee I.K. 1997. A simple method for RNA extraction from Marine macroalgae. Algae 12: 53-56.
  11. 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
  12. Lee T.K. and Lee H.K. 2003. A simple method for DNA extraction from red algae. Algae 18: 65-69 https://doi.org/10.4490/ALGAE.2003.18.1.065
  13. Manning K. 1991. Isolation of nucleic acids from plants by differential solvent precipitation. Anal. Biochem. 195: 45-50 https://doi.org/10.1016/0003-2697(91)90292-2
  14. Mayes C., Saunders G.W., Tan I.H. and Druehl L.D. 1992. DNA extraction methods for kelp (Laminariales) tissue. J. Phycol. 28: 714-716
  15. Mizukami Y., Okauchi M., Kito H., Ishimoto S.-I. Ishida T. and Fuseya M. 1995. Culture and development of electrically fused protoplasts from red algae, Porphyra yezoensis and P. suborbiculata. Aquaculture 132: 361-367 https://doi.org/10.1016/0044-8486(95)93662-T
  16. Mizukami Y., Kito H., Kunimoto M. and Kobayashi M. 1998. Effect of DNA preparation from laver (Porphyra yezoensis) thalli on reproducibility of RAPD (random amplified polymorphic DNA) patterns. J. Appl. Phycol. 10: 23-29 https://doi.org/10.1023/A:1008023806500
  17. Nakajima M., Kitade Y., Iitsuka O., Fukuda S. and Saga N. 2000. Rapid extraction of high-quality genomic DNA from Porphyra yezoensis (Bangiales, Rhodophyta). Phycological Research 48: 15-17
  18. Pandey R.N., Adams R.P. and Flournoy L.E. 1996. Inhibition of random amplified polymorphic DNAs(RAPDs) by plant polysaccharides. Plant Mol. Biol. Reporter 14: 17-22 https://doi.org/10.1007/BF02671898
  19. Parrish K.D. and Greenberg E.P. 1995. A Rapid Method for extraction and purification of DNA from Dental Plaque. Appl. Environ. Microbiol. 61: 4120-4123
  20. Pikaart M.J. and Villeponteau B. 1993. Suppression of PCR amplification by high levels of RNA. Bio Techniques 14: 14-25
  21. Ragan M.A. 1981 Chemical constituents of seaweeds. In: Lobban C. S. and Wynne M. J. (eds), The biology of seaweeds. University of California Press, Berkeley, pp. 25-35
  22. Roell M.K. and Morse D.E. 1991. Fractionation of nuclear, chloroplast, and mitochondrial DNA from Polysiphonia boldii (Rhodophyta) using a rapid and simple method for simultaneous isolation of RNA and DNA. J. Phycol. 27: 299- 305 https://doi.org/10.1111/j.0022-3646.1991.00299.x
  23. Xing S. 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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