4.5 Article

IMPROVED METHODS FOR THE ISOLATION OF CYANOBACTERIAL DNA FROM ENVIRONMENTAL SAMPLES

Journal

JOURNAL OF PHYCOLOGY
Volume 45, Issue 2, Pages 517-521

Publisher

WILEY-BLACKWELL PUBLISHING, INC
DOI: 10.1111/j.1529-8817.2009.00651.x

Keywords

cyanobacteria; DNA; fluorometry; humics; lake water; microcystin; PEG; PVPP; sediment; xanthogenate

Funding

  1. St. Johns River Water Management District

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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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