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

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.

authors

• Pillai, Suresh

published proceedings

• Appl Environ Microbiol

altmetric score

• 3

author list (cited authors)

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

citation count

• 159

complete list of authors

• Pillai, SD||Josephson, KL||Bailey, RL||Gerba, CP||Pepper, IL

publication date

• August 1991

publisher

• American Society for Microbiology  Publisher

published in

• Applied and Environmental Microbiology  Journal

keywords

• Base Sequence
• Centrifugation, Density Gradient
• DNA Transposable Elements
• DNA, Bacterial
• Electrophoresis, Agar Gel
• Molecular Sequence Data
• Polymerase Chain Reaction
• Sensitivity And Specificity
• Soil Microbiology

PubMed Central ID

• 1662931

Digital Object Identifier (DOI)

• 10.1128/aem.57.8.2283-2286.1991

start page

• 2283

end page

• 2286

volume

• 57

issue

• 8

URL

• http://dx.doi.org/10.1128/aem.57.8.2283-2286.1991