An improved method for efficient isolation and purification of genomic DNA from filamentous cyanobacteria belonging to genera Anabaena, Nodularia and Nostoc

Abstract

The aim of this work was to develop an improved method for isolation and purification of genomic DNA from filamentous cyanobacteria. The method described here employs a modified phenol extraction-based procedure. It allowed us to obtain a high yield (60–620 μg/g wet weight, depending on the cyanobacterial strain) of pure and undegraded genomic DNA (A260/A280 ratio of about 1.8 and A260/A230 ratio of about 2.0). Genomic DNA, isolated from cyanobacteria belonging to the genera Anabaena, Nodularia and Nostoc has been successfully used for construction of gene libraries. Thus, this method can be used in procedures requiring highly purified cyanobacterial DNA.

This is a preview of subscription content, access via your institution.

References

  1. Bertozzini, E., Penna, A., Pierboni, E., Bruce, I.J. and Magnani, M. (2005) Development of new procedures for the isolation of phytoplankton DNA from fixed samples. J Appl Phycol 17, 223–229.

    Article  Google Scholar 

  2. Bollet, C., Gevaudan, M.J., de Lamballerie, X., Zandotti, C. and de Micco, P. (1991) A simple method for the isolation of chromosomal DNA from gram positive or acid-fast bacteria. Nucleic Acids Res 19, 1955.

    Article  Google Scholar 

  3. Burja, A.M., Banaigs, B., Abou-Mansour, E., Burgess, J.G. and Wright, P.C. (2001) Marine cyanobacteria — a prolific source of natural products. Tetrahedron 57, 9347–9377.

    Article  Google Scholar 

  4. Fiore, M.F., Moon, D.H., Tsai, S.M., Lee, H. and Trevors, J.T. (2000) Miniprep DNA isolation from unicellular and filamentous cyanobacteria. J Microbiol Meth 39, 159–169.

    Article  Google Scholar 

  5. Hoiczyk, E. and Hansel, A. (2000) Cyanobacterial cell walls: news from an unusual prokaryotic envelope. J Bacteriol 182, 1191–1199.

    Article  Google Scholar 

  6. Kennedy, J., Flemer, B., Jackson, S.A., Lejon, D.P.H., Morrissey, J.P., O’Gara, F. and Dobson, A.D.W. (2010) Marine metagenomics: new tools for the study and exploitation of marine microbial metabolism. Mar Drugs 8, 608–628.

    Article  Google Scholar 

  7. Liberton, M. and Pakrasi, H.B. (2008) Membrane systems in cyanobacteria. In: The cyanobacteria: molecular biology, genomics and evolution eds. Herrero, A. and Flores, E., pp 277–285, Norwich: Caister Academic Press.

    Google Scholar 

  8. Manchester, K.L. (1995) Value of A260/A280 ratios for measurement of purity of nucleic acids. Biotechniques 19, 208–210.

    Google Scholar 

  9. Morin, N., Vallaeys, T., Hendrickx, L., Natalie, L. and Wilmotte, A. (2010) An efficient DNA isolation protocol for filamentous cyanobacteria of the genus Arthrospira. J Microbiol Meth 80, 148–154.

    Article  Google Scholar 

  10. Pope, P.B. and Patel, B.K.C. (2008) Metagenomic analysis of a freshwater toxic cyanobacteria bloom. FEMS Microbiol Ecol 64, 9–27.

    Article  Google Scholar 

  11. Saha, S.K., Uma, L. and Subramanian, G. (2005) An improved method for marine cyanobacterial DNA isolation. World J Microbiol Biotechnol 21, 877–881.

    Article  Google Scholar 

  12. Sambrook, J. and Russell, D.W. (2001) Molecular cloning: a laboratory manual. 3rd edition. Cold Spring Harbor (NY): Cold Spring Harbor Laboratory Press.

    Google Scholar 

  13. Sinha, S., Dadheech, P.K., KrishnaMo, M. (2009) Rapid DNA extraction from cyanobacteria. www.protocol-online.org. Date added: Feb 02, 2009

    Google Scholar 

  14. Sivonen, K. and Börner, T. (2008) Bioactive compounds produced by cyanobacteria. In: The cyanobacteria: molecular biology, genomics and evolution, eds. Herrero, A. and Flores, E. pp. 159–183, Norwich: Caister Academic Press.

    Google Scholar 

  15. Sivonen, K., Leikoski, N., Fewer, D.P. and Jokela, J. (2010) Cyanobactins—ribosomal cyclic peptides produced by cyanobacteria. Appl Microbiol Biotechnol 86, 1213–1225.

    Article  Google Scholar 

  16. Wu, X., Zarka, A. and Boussiba, S. (2000) A simplified protocol for preparing DNA from filamentous cyanobacteria. Plant Mol Biol Rep 18, 385–392.

    Article  Google Scholar 

Download references

Author information

Affiliations

Authors

Corresponding author

Correspondence to Grzegorz Węgrzyn.

About this article

Cite this article

Kaczyńska, A., Łoś, M. & Węgrzyn, G. An improved method for efficient isolation and purification of genomic DNA from filamentous cyanobacteria belonging to genera Anabaena, Nodularia and Nostoc . Ocean and Hydro 42, 8–13 (2013). https://doi.org/10.2478/s13545-013-0058-y

Download citation

Key words

  • DNA isolation
  • DNA purification
  • filamentous cyanobacteria
  • genomic DNA
  • genome library construction