UCRL-AR-145380 Ch. 4
Chapter 4: Effect of Ethanol on Hydrocarbon-
degrading Bacteria in the Saturated Zone:
Microbial Ecology Studies
Authors
Harry R. Beller*
Staci R. Kane*
Tina C. Legler*
October 2001
*Lawrence Livermore National Laboratory, Livermore, CA 94550
University of California • Livermore, CA 94550
Lawrence Livermore National Laboratory
Environmental Restoration Division
This work was performed under the auspices of the U. S. Department of Energy by University of California
Lawrence Livermore National Laboratory under Contract W-7405-Eng-48.
UCRL-AR-145380 Ch. 4
Subsurface Fate and Transport of Gasoline Containing Ethanol
October 2001
10/01/Ethanol Ch.4:HRB:rtd
4-iii
Table of Contents
Summary ..............................................................................................................................Summ-1
4. Effect of Ethanol on Hydrocarbon-degrading Bacteria in the Saturated Zone:
Microbial Ecology Studies..................................................................................................... 4-1
4.1. Introduction................................................................................................................... 4-1
4.2. Materials and Methods.................................................................................................. 4-3
4.2.1. Microcosm Construction and Sampling............................................................ 4-3
4.2.2. DNA Extraction ................................................................................................ 4-4
4.2.3. Real-Time, Quantitative Polymerase Chain Reaction (RTQ-PCR).................. 4-5
4.2.4. Data Analysis .................................................................................................... 4-6
4.2.5. Analyses of Toluene and Ethanol ..................................................................... 4-7
4.3. Results........................................................................................................................... 4-7
4.3.1. Performance of RTQ-