IMPROVED METHODS FOR THE ISOLATION OF CYANOBACTERIAL DNA FROM ENVIRONMENTAL SAMPLES1

Authors

  • Mete Yilmaz,

    1. Program in Fisheries and Aquatic Sciences, School of Forest Resources and Conservation, University of Florida, 7922 N.W. 71st Street, Gainesville, Florida 32653, USA
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  • Edward J. Phlips,

    1. Program in Fisheries and Aquatic Sciences, School of Forest Resources and Conservation, University of Florida, 7922 N.W. 71st Street, Gainesville, Florida 32653, USA
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  • Daniel Tillett

    1. School of Pharmacy and Applied Sciences, La Trobe University, P.O. Box 199, Bendigo, Victoria 3550, Australia
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  • 1

    Received 30 April 2008. Accepted 21 November 2008.

Abstract

DNA isolated from environmental samples often contains enzyme inhibitors disruptive to downstream molecular applications. Most of the existing methods of cyanobacterial DNA isolation do not effectively eliminate these inhibitors from sediment samples or cells collected from freshwater ecosystems. We describe improved methods based on the xanthogenate-SDS nucleic acid isolation (XS) method of Tillett and Neilan (2000). Our improved methods provided high-quality cyanobacterial DNA that could be amplified in PCR and digested with a restriction enzyme. Results were superior to several commercial kits. The DNA yield was also similar to that obtained via the standard XS method. These methods should provide valuable new tools for the expanded application of molecular genetics to limnological and oceanographic research.

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