This Article
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Jacobsen, C. S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Jacobsen, C. S.
Agricola
Right arrow Articles by Jacobsen, C. S.

 Previous Article  |  Next Article 

Appl. Environ. Microbiol., 09 1995, 3347-3352, Vol 61, No. 9
Copyright © 1995, American Society for Microbiology

Microscale detection of specific bacterial DNA in soil with a magnetic capture-hybridization and PCR amplification assay

CS Jacobsen
Department of Ecology and Molecular Biology, Royal Veterinary and Agricultural University, Frederiksberg C, Copenhagen, Denmark.

A magnetic capture-hybridization PCR technique (MCH-PCR) was developed to eliminate the inhibitory effect of humic acids and other contaminants in PCRs targeting specific soil DNA. A single-stranded DNA probe, which was complementary to an internal part of the target gene, was used to coat magnetic beads. After hybridization in a suspension of soil DNA, magnetic extraction of the beads separated the hybrid DNA from all other soil DNA, humic acids, and other interfering soil components. The MCH was followed by PCR amplification of the specific target DNA. In barley rhizosphere soil, detection of a lux gene inserted in a Pseudomonas fluorescens strain could be demonstrated in nonsterile soil samples (0.5 mg). This corresponded to a detection of fewer than 40 bacterial cells per cm of barley root. The MCH-PCR technique greatly improves the current protocols for PCR detection of specific microorganisms or genes in soil because specific target DNA sequences from very small soil samples can be extracted and determined.


This article has been cited by other articles:

  • de Vries, J., Heine, M., Harms, K., Wackernagel, W. (2003). Spread of Recombinant DNA by Roots and Pollen of Transgenic Potato Plants, Identified by Highly Specific Biomonitoring Using Natural Transformation of an Acinetobacter sp.. Appl. Environ. Microbiol. 69: 4455-4462 [Abstract] [Full Text]  
  • Chandler, D. P., Newton, G. J., Small, J. A., Daly, D. S. (2003). Sequence versus Structure for the Direct Detection of 16S rRNA on Planar Oligonucleotide Microarrays. Appl. Environ. Microbiol. 69: 2950-2958 [Abstract] [Full Text]  
  • Fredslund, L., Ekelund, F., Jacobsen, C. S., Johnsen, K. (2001). Development and Application of a Most-Probable-Number-PCR Assay To Quantify Flagellate Populations in Soil Samples. Appl. Environ. Microbiol. 67: 1613-1618 [Abstract] [Full Text]  
  • Spiro, A., Lowe, M., Brown, D. (2000). A Bead-Based Method for Multiplexed Identification and Quantitation of DNA Sequences Using Flow Cytometry. Appl. Environ. Microbiol. 66: 4258-4265 [Abstract] [Full Text]  
  • Chandler, D. P., Stults, J. R., Cebula, S., Schuck, B. L., Weaver, D. W., Anderson, K. K., Egholm, M., Brockman, F. J. (2000). Affinity Purification of DNA and RNA from Environmental Samples with Peptide Nucleic Acid Clamps. Appl. Environ. Microbiol. 66: 3438-3445 [Abstract] [Full Text]  
  • de Moraes, R. R., Maruniak, J. E., Funderburk, J. E. (1999). Methods for Detection of Anticarsia gemmatalis Nucleopolyhedrovirus DNA in Soil. Appl. Environ. Microbiol. 65: 2307-2311 [Abstract] [Full Text]  
  • Johnsen, K., Enger, O., Jacobsen, C. S., Thirup, L., Torsvik, V. (1999). Quantitative Selective PCR of 16S Ribosomal DNA Correlates Well with Selective Agar Plating in Describing Population Dynamics of Indigenous Pseudomonas spp. in Soil Hot Spots. Appl. Environ. Microbiol. 65: 1786-1788 [Abstract] [Full Text]  
  • Hauben, L., Steenackers, M., Swings, J. (1998). PCR-Based Detection of the Causal Agent of Watermark Disease in Willows (Salix spp.). Appl. Environ. Microbiol. 64: 3966-3971 [Abstract] [Full Text]  
  • Chen, J., Johnson, R., Griffiths, M. (1998). Detection of Verotoxigenic Escherichia coli by Magnetic Capture-Hybridization PCR. Appl. Environ. Microbiol. 64: 147-152 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»