Sampling Techniques

Methods and Methodology

Environmental Microbiology

BIOL/CEE/CSES/ENSC 4164

The students in the 1999 and 2000 classes prepared a webpage on one of the methods listed below as their individual class assignment. Gene Probes (under Detection of Microbial Populations - see below) was done by the instructor as an example to follow. The methods in black have not yet done. As this material is now eight years old, it's probably time to consider an upgrade to some of the methods, plus adding new ones that have been developed in recent years.

Sampling

Sample Collection and Storage

Sample Processing

Detection of Microbial Populations

Direct Isolation	Plant Trapping
Enrichment Cultures	Immunoassays and ELISA
Polymerase Chain Reaction (PCR)	Gene Probes
Bioluminescence	Percent G+C
The BIOLOG Assay	Electron Microscopy - 1
Luminometry

Determination of Microbial Numbers

Light Microscopy	Fluorescence Microscopy
Electron Microscopy - 2	Particle Counters
Most-Probable-Number Counts	Colony Hybridization
Direct Plate Counts	Fluorescence in situ Hybridization

Determination of Microbial Biomass

ATP and Total Adylate	Cell Wall Components
Chlorophyll and Pigments	Nucleic Acids
Fatty Acid Analysis

Measure of Microbial Metabolism

Respiration Rates	Enzymatic Assays
Biochemical Oxygen Demand (BOD)	Nucleotide Uptake

Development of Transgenic Microorganisms

Genomic Cloning	Preparation and Characterization of RNA
Preparation of cDNA and the Generation of cDNA libraries	Insertion of Foreign Genes and Vectors
Identification and Characterization of Specific Clones	Restriction Enzymes
Isolating and Characterizing Nucleic Acids