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

Kaczyńska, Angelika; Łoś, Marcin; Węgrzyn, Grzegorz

doi:10.2478/s13545-013-0058-y

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.

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. http://dx.doi.org/10.1007/s10811-005-2130-510.1007/s10811-005-2130-5
Search in 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. http://dx.doi.org/10.1093/nar/19.8.195510.1093/nar/19.8.1955
Search in 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. http://dx.doi.org/10.1016/S0040-4020(01)00931-010.1016/S0040-4020(01)00931-0
Search in 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. http://dx.doi.org/10.1016/S0167-7012(99)00110-410.1016/S0167-7012(99)00110-4
Search in Google Scholar
[5] Hoiczyk, E. and Hansel, A. (2000) Cyanobacterial cell walls: news from an unusual prokaryotic envelope. J Bacteriol 182, 1191–1199. http://dx.doi.org/10.1128/JB.182.5.1191-1199.200010.1128/JB.182.5.1191-1199.20009440210671437
Search in 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. http://dx.doi.org/10.3390/md803060810.3390/md8030608285735420411118
Search in 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.
Search in Google Scholar
[8] Manchester, K.L. (1995) Value of A260/A280 ratios for measurement of purity of nucleic acids. Biotechniques 19, 208–210.
Search in 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. http://dx.doi.org/10.1016/j.mimet.2009.11.01210.1016/j.mimet.2009.11.01220004220
Search in 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. http://dx.doi.org/10.1111/j.1574-6941.2008.00448.x10.1111/j.1574-6941.2008.00448.x18328084
Search in 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. http://dx.doi.org/10.1007/s11274-004-6187-010.1007/s11274-004-6187-0
Search in 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.
Search in 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
Search in 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.
Search in 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. http://dx.doi.org/10.1007/s00253-010-2482-x10.1007/s00253-010-2482-x285435320195859
Search in 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. http://dx.doi.org/10.1007/BF0282506710.1007/BF02825067
Search in Google Scholar

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

Abstract

Paradigm

My account