NDLI: Extraction from Natural Planktonic Microorganisms of DNA Suitable for Molecular Biological Studies

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Populations of Anaerobic Phototrophic Bacteria in a Spartina alterniflora Salt Marsh

Plugging of a Model Rock System by Using Starved Bacteria

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Evidence for plasmid-mediated toxin and bacteriocin production in Clostridium botulinum type G.

Sub-Parts-Per-Billion Nitrate Method: Use of an N2O-Producing Denitrifier to Convert NO3 − or 15NO3 − to N2O †

Bacterial metabolism of carbofuran.

The mannoprotein of Saccharomyces cerevisiae is an effective bioemulsifier.

Extraction from Natural Planktonic Microorganisms of DNA Suitable for Molecular Biological Studies

Sources of Variability in the Measurement of Fungal Spore Yields

Bioluminescence Assay for Measuring the Number of Bacteria Adhering to the Hydrocarbon Phase in the BATH Test

Microbially Mediated Mn(II) Oxidation in an Oligotrophic Arctic Lake †

Factors influencing Clostridium botulinum spore germination, outgrowth, and toxin formation in acidified media.

Evaluation of selective direct plating media for their suitability to recover uninjured, heat-injured, and freeze-injured Listeria monocytogenes from foods.

Hydrogen Partial Pressures in a Thermophilic Acetate-Oxidizing Methanogenic Coculture

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Isolation and Preliminary Characterization of Twenty Bacteriophages Infecting Either Brevibacterium or Arthrobacter Strains

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Purification and Comparison of the Periplasmic and Extracellular Forms of the Cellodextrinase from Bacteroides succinogenes

Reliability of the Most-Probable-Number Technique for Enumerating Rhizobia in Tropical Soils †

Toluene degradation by Pseudomonas putida F1: genetic organization of the tod operon.

Growth of and fumitremorgin production by Neosartorya fischeri as affected by temperature, light, and water activity.

Cloning of the cellulase gene from Ruminococcus albus and its expression in Escherichia coli.

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Plasmid profiles as indicators of the source of contamination of Staphylococcus aureus endemic within poultry processing plants.

Cloning of Trametes versicolor Genes Induced by Nitrogen Starvation

Aerial Dispersal and Epiphytic Survival of Pseudomonas syringae during a Pretest for the Release of Genetically Engineered Strains into the Environment

Monoclonal antibodies which identify a genus-specific Listeria antigen.

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Isolation and characterization of a temperate bacteriophage from the ruminal anaerobe Selenomonas ruminantium.

l-Methionine, a Precursor of Trace Methane in Some Proteolytic Clostridia

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Plasmid profile analysis of a salmonellosis outbreak and identification of a restriction and modification system.

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Molecular filtration for recovery of waterborne viruses of fish.

Genetic Transformation of an argB Mutant of Aspergillus oryzae

Mineralization of polycyclic aromatic hydrocarbons by a bacterium isolated from sediment below an oil field.

Formation and regeneration of protoplasts and spheroplasts of gastrointestinal strains of lactobacilli.

Degradation of DNA in cells and extracts of the obligately anaerobic bacterium Roseburia cecicola upon exposure to air.

Presence of aflatoxin M1 in commercial ultra-high-temperature-treated milk.

Synthesis of Exopolysaccharide by Bradyrhizobium japonicum during Growth on Hydroaromatic Substrates

A pH Control System Based on Malate Decarboxylation for the Cultivation of Lactic Acid Bacteria †

A Simple, Rapid Method for Enumerating Total Viable and Metabolically Active Bacteria in Groundwater

Incidence of coliphage in potable water supplies.

Methylcellulose inhibition of exo-beta-1,4-glucanase A from Ruminococcus flavefaciens FD-1.

Morphological diversity of ruminal bacteriophages from sheep and cattle.

Trichloroethylene Metabolism by Microorganisms That Degrade Aromatic Compounds

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Extraction from Natural Planktonic Microorganisms of DNA Suitable for Molecular Biological Studies

Content Provider	PubMed Central
Author	Fuhrman, Jed A. Comeau, Dorothy E. Hagström, Åke Chan, Amy M.
Abstract	We developed a simple technique for the high-yield extraction of purified DNA from mixed populations of natural planktonic marine microbes (primarily bacteria). This is a necessary step for several molecular biological approaches to the study of microbial communities in nature. The microorganisms from near-shore marine and brackish water samples, ranging in volume from 8 to 40 liters, were collected on 0.22-μm-pore-size fluorocarbon-based filters, after prefiltration through glass fiber filters, to remove most of the eucaryotes. DNA was extracted directly from the filters in 1% sodium dodecyl sulfate that was heated to 95 to 100°C for 1.5 to 2 min. This procedure lysed essentially all the bacteria and did not significantly denature the DNA. The DNA was purified by phenol extraction, and precautions were taken to minimize shearing. Agarose gel electrophoresis showed that most of the final preparation had a large molecular size (>23 kilobase pairs). The DNA was sufficiently pure to allow complete digestion by the restriction endonuclease Sau3AI and ligation to vector DNA. In a sample in which the extracted DNA was quantified by binding to the dye Hoechst H33258, DNA was quantitatively extracted, and 45% of the initially extracted DNA was recovered after purification. Final yields were a few micrograms of DNA per liter of seawater and were roughly 25 to 50% of the total bacterial DNA in the sample. Alternatives to the initial harvest by filtration method, including continuous-flow centrifugation and thin-channel or hollow-fiber concentration followed by centrifugation, were less efficient than filtration in terms of both time and yield, largely because of the difficulty of centrifuging the very small bacteria typical of marine plankton. These methods were judged to be less appropriate for studies of natural populations as they impose a strong selection for the larger bacteria.
Starting Page	1426
File Format	PDF
ISSN	10985336
e-ISSN	10985336
Journal	Applied and Environmental Microbiology
Issue Number	6
Volume Number	54
Language	English
Publisher Date	1988-06-01
Access Restriction	Open
Subject Keyword	Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Ecology Food Science Applied Microbiology and Biotechnology Biotechnology

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