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Applied and Environmental Microbiology, May 2008, p. 3302-3305, Vol. 74, No. 10
0099-2240/08/$08.00+0     doi:10.1128/AEM.02630-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Recovery, Purification, and Cloning of High-Molecular-Weight DNA from Soil Microorganisms{triangledown}

Mark R. Liles,1 Lynn L. Williamson,2 Jitsupang Rodbumrer,3 Vigdis Torsvik,4 Robert M. Goodman,5* and Jo Handelsman2

Department of Biological Sciences, Auburn University, Auburn, Alabama,1 Department of Bacteriology, University of Wisconsin, Madison, Wisconsin,2 Department of Biotechnology, Mahidol University, Bangkok, Thailand,3 Department of Microbiology, University of Bergen, Bergen, Norway,4 School of Environmental and Biological Sciences, Rutgers, The State University of New Jersey, New Brunswick, New Jersey5

Received 20 November 2007/ Accepted 12 March 2008

We describe here an improved method for isolating, purifying, and cloning DNA from diverse soil microbiota. Soil microorganisms were extracted from soils and embedded and lysed within an agarose plug. Nucleases that copurified with the metagenomic DNA were removed by incubating plugs with a high-salt and -formamide solution. This method was used to construct large-insert soil metagenomic libraries.

* Corresponding author. Mailing address: Rutgers, The State University of New Jersey, School of Environmental and Biological Sciences, 88 Lipman Drive, Suite 104, New Brunswick, NJ 08901. Phone: (732) 932-9000, ext. 501. Fax: (732) 932-8955. E-mail: rgoodman{at}aesop.rutgers.edu

{triangledown} Published ahead of print on 21 March 2008.

Applied and Environmental Microbiology, May 2008, p. 3302-3305, Vol. 74, No. 10
0099-2240/08/$08.00+0     doi:10.1128/AEM.02630-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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