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MALDI-TOF Analysis of Polymerase Chain Reaction Products from Methanotrophic Bacteria

Hurst, Gregory B. and Weaver, Kristal and Doktycz, Mitchel J. and Buchanan, Michelle V. and Costello, Andria M. and Lidstrom, Mary E. (1998) MALDI-TOF Analysis of Polymerase Chain Reaction Products from Methanotrophic Bacteria. Analytical Chemistry, 70 (13). pp. 2693-2698. ISSN 0003-2700. doi:10.1021/ac980044e.

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Polymerase chain reaction (PCR) assays were designed to amplify 56- and 99-base regions of the pmoA gene from Methylosinus trichosporium OB3b and Methylomicrobium albus BG8, two species of methanotrophic bacteria that are of interest for monitoring bioremediation activity. The PCR product sizes are in a mass range that is accessible to analysis by MALDI-TOF mass spectrometry. A rapid purification procedure using commercially available reversed-phase cartridges was applied prior to MALDI-TOF analysis. A small aliquot (1.5%, 1.5 μL) from a single 100-μL PCR reaction was sufficient for reliable detection. No cross-amplification products were observed when primers designed for one bacterial species were used with genomic DNA of the other species. The methodology described here has potential to allow less expensive and faster characterization of the ability of microbial populations to destroy pollutants in groundwater and soil at contaminated industrial sites.

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Additional Information:© 1998 American Chemical Society. Received for review January 13, 1998. Accepted March 31, 1998. Publication Date (Web): May 5, 1998. K.W. acknowledges support through an appointment to the Oak Ridge National Laboratory Postgraduate Research Program, administered jointly by the Oak Ridge Institute for Science and Education and Oak Ridge National Laboratory. This research was sponsored by the U.S. Department of Energy, Environmental Management Science Program and Office of Biological and Environmental Research, under contract no. DE-AC05-96OR22464 with Oak Ridge National Laboratory, managed by Lockheed Martin Energy Research Corp.
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Oak Ridge National LaboratoryUNSPECIFIED
Department of Energy (DOE)DE-AC05-96OR22464
Issue or Number:13
Record Number:CaltechAUTHORS:20171120-133844701
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Official Citation:MALDI-TOF Analysis of Polymerase Chain Reaction Products from Methanotrophic Bacteria. Gregory B. Hurst, Kristal Weaver, Mitchel J. Doktycz, Michelle V. Buchanan, Andria M. Costello, and Mary E. Lidstrom. Analytical Chemistry 1998 70 (13), 2693-2698 DOI: 10.1021/ac980044e
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:83348
Deposited By: Tony Diaz
Deposited On:20 Nov 2017 21:48
Last Modified:15 Nov 2021 19:57

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