Animal cells and systems

The Korean Society for Integrative Biology (한국통합생물학회)
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Efficient Isolation of Intact RNA from the Soft Coral Scleronephthya gracillimum $(K\ddot{u}kenthal)$ for Gene Expression Analyses

  • Woo, Seo-Nock (Southern Coastal Environment Research Division, Korea Ocean Research and Development Institute) ;
  • Yum, Seung-Shic (Southern Coastal Environment Research Division, Korea Ocean Research and Development Institute) ;
  • Yoon, Moon-Geun (Department of Biological Science, Sungkyunkwan University) ;
  • Kim, Sa-Heung (Laboratory of Marine Biodiversity, In The Sea Korea Co., Ltd.) ;
  • Lee, Jong-Rak (Laboratory of Marine Biodiversity, In The Sea Korea Co., Ltd.) ;
  • Kim, Jeong-Ha (Department of Biological Science, Sungkyunkwan University) ;
  • Lee, Taek-Kyun (Southern Coastal Environment Research Division, Korea Ocean Research and Development Institute)
  • Published : 2005.12.01
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Abstract

A rapid, simple and efficient method to extract RNA from the adult polyps of a soft coral, marine cnidarian, Scleronephthya gracillimum $(K\ddot{u}kenthal)$; was developed in this study. The highest yield and purity of RNA was obtained with the lysis solution containing 35 mM EDTA, 0.7 M LiCl, 7.0% SDS, and 200 mM Tris-Cl (pH 9.0). Approximately $40{\mu}g$ of total RNA was extracted from 200 mg of liquid nitrogen-pulverized polyp tissue. The ratio of absorbance at 260 nm and 280 nm ranged from 1.8 to 2.0. The results of the reverse transcription polymerase chain reaction (RTPCR) with ${\beta}-actin$ gene specific primers and Northern blot analysis using the same gene probe revealed that the RNA extracted by our method had high quality, and was sufficient for subsequent molecular biological analyses. This method was effective for RNA extraction from other soft coral species which belong to the genus Dendronephthya.

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References

  1. Chirgwin JM, Przybyla AE, MacDonald RJ, and Rutter WJ (1979) Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18: 5294-5299 https://doi.org/10.1021/bi00591a005
  2. Chomczynski P (1993) A reagent for the single-step simultaneous isolation of RNA, DNA and proteins from cell and tissue samples. BioTechniques 15: 532-537
  3. Chomczynski P and Mackey M (1995) Substitution of chloroform by bromochloropropane in the single-step method of RNA isolation. Anal Biochem 225: 163-164 https://doi.org/10.1006/abio.1995.1126
  4. Chomczynski P and Sacchi N (1987) Single-step method of RNA isolation by acid guanidium thiocyanate-phenol-chloroform extraction. Anal Biochem 161: 156-159 https://doi.org/10.1016/0003-2697(87)90021-2
  5. Fabricius K and Alderslade P (2001) Soft Corals and Sea Fans. New Litho, Melbourne. pp 1-264
  6. Groppe JC and Morse DE (1993) Isolation of full-length RNA templates for reverse transcription from tissues rich in RNase and proteoglycans. Analy Biochem 210: 337-343 https://doi.org/10.1006/abio.1993.1205
  7. Hipfel R, Garbe C, and Schittek B (1998) RNA isolation from human skin tissues for colorimetric differential display. J Biochem Biophys Meth 37: 131-135 https://doi.org/10.1016/S0165-022X(98)00023-2
  8. Lee Y-J and Song J-I (2000) Phylogenetic analysis of the genus Dendronephthya (Nephtheidae, Alcyonacea) based on internal transcribed spacer sequences of nuclear DNA. Korean J Biol Sci 4: 319-324 https://doi.org/10.1080/12265071.2000.9647563
  9. Michel U, Rau A and Rieckmann P (1997) Isolation of cytoplasmic RNA from small samples of whole blood. Anal Biochem 249: 246-247 https://doi.org/10.1006/abio.1997.2174
  10. Mukhtar M, Parveen Z, and Logan DA (1998) Isolation of RNA from the filamentous fungus Mucor circinelloides. J Microbiol Meth 33: 115-118 https://doi.org/10.1016/S0167-7012(98)00047-5
  11. Nuyts S, Mellaert LV, and Anne PL (2001). Efficient isolation of total RNA from Clostrium without DNA contamination. J Microbiol Meth 44: 235-238 https://doi.org/10.1016/S0167-7012(01)00219-6
  12. Sambrook J and Russell DW (2001) Molecular Cloning: a Laboratory Manual. 3rd Ed. Cold Spring Harbor Laboratory Press, New York
  13. Seo Y, Jung JH, Rho JR, Shin J, and Song J-I (1995) Isolation of novel bioactive steroids from the soft coral Alcyonium gracillimum. Tetrahedron 51: 2497-2506 https://doi.org/10.1016/0040-4020(95)00014-Y
  14. Tavangar K, Hoffman AR, and Kraemer F (1990) A micromethod for the isolation of total RNA from adipose tissue. Anal Biochem 186: 60-63 https://doi.org/10.1016/0003-2697(90)90572-Q
  15. Vicient CM and Delseny M (1999) Isolation of total RNA from Arabidopsis thaliana seeds. Anal Biochem 268: 412-413 https://doi.org/10.1006/abio.1998.3045
  16. Warner SAJ (1996) Genomic DNA isolation and lambda library construction. In: Foster GD and Twell D (eds), Plant Gene Isolation. Principles and Practice, John Wiley & Sons, Chichester, pp 51-74
  17. Whitchel HJ, Maitland NJ, and Meech RW (1996) Milligram quantity preparation of RNA from a marine invertebrate with a high fluid content. BioTechniques 21: 1024-1026