Rapid method for processing soil samples for polymerase chain reaction amplification of specific gene sequences

Author:

Pillai S D1,Josephson K L1,Bailey R L1,Gerba C P1,Pepper I L1

Affiliation:

1. Department of Soil and Water Science, University of Arizona, Tucson 85721.

Abstract

Bacterial cells can be differentially separated from soil colloids on the basis of their buoyant densities. By using this principle, a modified sucrose gradient centrifugation protocol has been developed for separating bacterial cells from most of the soil colloids. Since the bacterial cell suspension still contained some colloidal soil particles, which inhibited polymerase chain reaction amplification, a new "double" polymerase chain reaction method of analysis was adopted for amplification of Tn5-specific gene sequences. This new protocol allowed rapid detection of small numbers (1 to 10 CFU/g) of bacterial cells present in soil samples.

Publisher

American Society for Microbiology

Subject

Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology

Reference16 articles.

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4. Regulation of of Tn 5 transposition in Salmonella typhimurium;Biek D.;Proc. Natl. Acad. Sci. USA,1980

5. Enumeration of TnS mutant bacteria in soil by mostprobable-number DNA hybridization procedure and antibiotic resistance;Frederickson J. K.;Appl. Environ. Microbiol.,1988

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