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Applied and Environmental Microbiology, August 2005, p. 4893-4896, Vol. 71, No. 8
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.8.4893-4896.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

SHORT REPORT

Fluorophore-Labeled Primers Improve the Sensitivity, Versatility, and Normalization of Denaturing Gradient Gel Electrophoresis

Josh D. Neufeld and William W. Mohn*

Department of Microbiology and Immunology, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada

Received 22 November 2004/ Accepted 28 February 2005

Denaturing gradient gel electrophoresis (DGGE) is widely used in microbial ecology. We tested the effect of fluorophore-labeled primers on DGGE band migration, sensitivity, and normalization. The fluorophores Cy5 and Cy3 did not visibly alter DGGE fingerprints; however, 6-carboxyfluorescein retarded band migration. Fluorophore modification improved the sensitivity of DGGE fingerprint detection and facilitated normalization of samples from multiple gels by the application of intralane standards.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, University of British Columbia, 300-6174 University Boulevard, Vancouver, British Columbia V6T 1Z3, Canada. Phone: 604-822-4285. Fax: 604-822-6041. E-mail: wmohn{at}interchange.ubc.ca.

Applied and Environmental Microbiology, August 2005, p. 4893-4896, Vol. 71, No. 8
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.8.4893-4896.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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