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Applied and Environmental Microbiology, December 2000, p. 5488-5491, Vol. 66, No. 12
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Rapid Method for Coextraction of DNA and RNA from Natural Environments for Analysis of Ribosomal DNA- and rRNA-Based Microbial Community Composition

Robert I. Griffiths,1,2 Andrew S. Whiteley,1 Anthony G. O'Donnell,2 and Mark J. Bailey1,*

Molecular Microbial Ecology Laboratory, Institute of Virology and Environmental Microbiology, CEH-Oxford, Oxford OX1 3SR,1 and Department of Agricultural and Environmental Science, The University of Newcastle Upon Tyne, Newcastle Upon Tyne NE1 7RU,2 United Kingdom

Received 5 June 2000/Accepted 13 September 2000

A rapid protocol for the extraction of total nucleic acids from environmental samples is described. The method facilitates concomitant assessment of microbial 16S rRNA diversity by PCR and reverse transcription-PCR amplification from a single extraction. Denaturing gradient gel electrophoresis microbial community analysis differentiated the active component (rRNA derived) from the total bacterial diversity (ribosomal DNA derived) down the horizons of an established grassland soil.

* Corresponding author. Mailing address: IVEM, CEH-Oxford, Mansfield Rd., Oxford OX1 3SR, United Kingdom. Phone: 44 (0)1865 281630. Fax: 44 (0)1865 281696. E-mail: mbj{at}ceh.ac.uk.

Applied and Environmental Microbiology, December 2000, p. 5488-5491, Vol. 66, No. 12
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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