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An Improved RNA Isolation Method for Filamentous Fungus Blakeslea trispora Rich in Polysaccharides

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Abstract

Isolation and purification of biologically active RNA from filamentous fungi is difficult because of the complex cell wall and the high level of polysaccharides which bind to or co-precipitate with RNA. Using benzyl chloride and guanidine thiocyanate, RNA was successfully isolated from Blakeslea trispora in which other RNA extraction methods and commercially available kits failed to deliver suitable results. The RNA isolated by this procedure appears to be relatively pure, as it has a ratio of absorbance at 260/280 nm of 1.8–1.9. The integrity of the RNA was further substantiated by RT-PCR and Northern hybridization, respectively. This procedure should be useful for isolating RNA from other filamentous fungi and, therefore, will serve as an important tool for the molecular analysis of these organisms.

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References

  1. Lee, P. C., & Schmidt-Dannert, C. (2002). Applied Microbiology and Biotechnology, 60, 1–11 Medline. DOI 10.1007/s00253-002-1101-x.

    Article  CAS  Google Scholar 

  2. Jayaram, R. (1986). Proceedings of the National Academy of Sciences of the United States of America, 83, 7344–7347 Medline. DOI 10.1073/pnas.83.19.7344.

    Article  Google Scholar 

  3. Quiles-Rosillo, M. D., Ruiz-Vazquez, R. M., Torres-Martinez, S., & Garre, V. (2003). FEMS Microbiology Letters, 222, 229–236 Medline.

    Article  CAS  Google Scholar 

  4. Mukhtar, M., Parveen, Z., & Logana, D. A. (1998). Journal of Microbiological Methods, 33, 115–118 DOI 10.1016/S0167-7012(98)00047-5.

    Article  CAS  Google Scholar 

  5. Yang, H., Wang, S. C., & Wang, Z. T. (2004). Chin. Pharm. J., 39, 254–256.

    CAS  Google Scholar 

  6. Li, B., Wang, B., Tang, K., Liang, Y. L., Wang, J., & Wei, J. M. (2006). Colloids and Surfaces B: Biointerfaces, 49, 101–5 DOI 10.1016/j.colsurfb.2006.03.006..

    Article  CAS  Google Scholar 

  7. Chomczynski, P., & Sacchi, N. (1987). Analytical Biochemistry, 162, 156–159 Medline. DOI 10.1016/0003-2697(87)90021-2.

    Article  CAS  Google Scholar 

  8. Majumdar, D., Avissar, Y. J., & Wyche, J. H. (1991). BioTechniques, 11, 94–101 Medline.

    CAS  Google Scholar 

  9. Dessel, W. V., Mellaert, L. V., Geukens, N., Lammertyn, E., & Anne, J. (2004). Journal of Microbiological Methods, 58, 135–137 Medline. DOI 10.1016/j.mimet.2004.03.015.

    Article  Google Scholar 

  10. Keulen, G. V., Siebring, J., Rembacz, K. P., Hoogeveen, M., Tomczynska, , & Dijkhuizen, L. (2004). Journal of Microbiological Methods, 58, 139–142 Medline. DOI 10.1016/j.mimet.2004.03.014.

    Article  Google Scholar 

  11. Lopez-Gomez, R., & Gomez-Lim, M. A. (1992). HortScience, 27, 440–442.

    CAS  Google Scholar 

  12. Sharma, A. D., Gill, P. K., & Singh, P. (2003). Analytical Biochemistry, 314, 319–21 Medline. DOI 10.1016/S0003-2697(02)00689-9.

    Article  CAS  Google Scholar 

  13. Shan, Z. P., Meng, Y., & Jiang, W. H. (2001). Biotechnology (Chinese), 11, 5–7.

    Google Scholar 

  14. Chirgwin, J. M., Pryzbyla, A. E., & Macdonald, R. J. (1979). Biochemistry, 18, 5294–5299 Medline. DOI 10.1021/bi00591a005.

    Article  CAS  Google Scholar 

  15. Raina, K., & Chandlee, J. M. (1996). BioTechniques, 21, 1030–1032 Medline.

    CAS  Google Scholar 

Download references

Acknowledgements

The authors would like to acknowledge the financial support of the National Natural Science Foundation of China (No. 20376007), the Foundation of Key Laboratory of Bioprocess of Beijing (Grant No. SYS 100100421), the National Natural Science Foundation of China (Grant no. 20576010), the National Natural Science Foundation of China (No. 20776009), and the Program for New Century Excellent Talents (NCET-05-0117).

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Authors and Affiliations

  1. Key Laboratory of Bioprocess of Beijing, Beijing University of Chemical Technology, Beijing, 100029, China

    Ye Li, Wenya Wang, Xueling Du & Qipeng Yuan

  2. The College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, 100029, China

    Wenya Wang

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  1. Ye Li
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  2. Wenya Wang
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  3. Xueling Du
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  4. Qipeng Yuan
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Correspondence to Qipeng Yuan.

Additional information

Ye Li and Wenya Wang contributed to the work equally.

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Li, Y., Wang, W., Du, X. et al. An Improved RNA Isolation Method for Filamentous Fungus Blakeslea trispora Rich in Polysaccharides. Appl Biochem Biotechnol 160, 322–327 (2010). https://doi.org/10.1007/s12010-008-8289-x

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  • DOI: https://doi.org/10.1007/s12010-008-8289-x

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