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United States Department of Agriculture

Agricultural Research Service

Title: Monitoring Bacterial Gene Expression in Soil Using Polymerase Chain Reaction Amplification

Authors
item Perkins E J,
item Kennedy A C,

Submitted to: Agronomy Abstracts
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: November 1, 1995
Publication Date: N/A

Technical Abstract: Polymerase chain reaction (PCR) methods and DNA probe-based methods allow monitoring of soil microbial populations regardless of culturability. Soils may have high levels of total organic carbon that interfere with enzymatic reactions. Current DNA and RNA isolation procedures are difficult and time consuming, often requiring purification by gel filtration. We have developed a rapid protocol for isolation of both DNA and RNA from milligram quantities of soil. The method consists of a simple, one step microwave lysis. Using DNA isolated by this method, 16s ribosomal RNA genes were amplified from high carbon content soils and petroleum and herbicide contaminated soils. RNA isolated by this technique was suitable for reverse transcriptase-PCR. This protocol is useful in the isolation of 16s rRNA genes and transcripts for monitoring soil microbial diversity and gene expression.

Last Modified: 11/20/2014
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