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

S. D. Pillai, K. L. Josephson, R. L. Bailey, C. P. Gerba, I. L. Pepper

Research output: Contribution to journalArticlepeer-review

83 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)2283-2286
Number of pages4
JournalApplied and environmental microbiology
Volume57
Issue number8
DOIs
StatePublished - 1991

ASJC Scopus subject areas

  • Biotechnology
  • Food Science
  • Applied Microbiology and Biotechnology
  • Ecology

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