Detection and quantification of microbial DNA sequences in soil by Southern- and dot/slot blot hybridization

Carsten S. Jacobsen

Research output: Chapter in Book/Report/Conference proceedingChapter in bookResearchpeer-review

Abstract

Methods to extract high purity DNA directly from soil microorganisms are now easily available, allowing the application of direct hybridization techniques in soil
microbiology. Methods that can detect specific gene sequences, whether present in dead or functioning bacteria, or even as free DNA in the soil, have been developed. To achieve a sensitive detection, the choice has often been to use the polymerase chain reaction [9] to amplify the target gene in a separate step between DNA extraction and subsequent hybridization steps [12, 14]. The PCR approach is described in chapters 2.10, 2.11 and 3.1 in this manual, and a solution to solve the important problem of quantitative analysis using PCR is described in chapter 2.11, 2.13, 2.14 and 2.15.
Original languageEnglish
Title of host publicationMolecular microbial ecology manual (Second edition)
EditorsGeorge A. Kowalchuk, Frans J. de Bruijn, Ian M. Head, Antoon D.L. Akkermans, Jan Dirk van Elsas
Place of PublicationDordrecht, The Netherlands
PublisherKluwer Academic Publishers
Chapter2.08
Pages333-344
Number of pages12
Edition2
ISBN (Electronic)978-1-4020-2177-0
ISBN (Print)978-1-4020-2176-3
DOIs
Publication statusPublished - 2004

Programme Area

  • Programme Area 2: Water Resources

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