Soil conservation for DNA preservation for bacterial molecular studies

The aim of this study was to select a method for preserving bacterial DNA in soil samples in cases where there are no possibilities of using freezing or cooling methods. To overcome this difficulty, we hypothesized that adding absolute ethanol to soil samples could be as successful to preserve bacte...

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Bibliographic Details
Published in:European journal of soil biology 2000-01, Vol.36 (1), p.51-55
Main Authors: Harry, Myriam, Gambier, Béatrice, Garnier-Sillam, Evelyne
Format: Article
Language:English
Subjects:
ADN
Agronomy. Soil science and plant productions
BACTERIA
Bacteriology
Biochemistry and biology
BIOLOGIA MOLECULAR
Biological and medical sciences
BIOLOGIE MOLECULAIRE
Chemical, physicochemical, biochemical and biological properties
conservation
DNA
DNA recovery
Fundamental and applied biological sciences. Psychology
Genetics
Microbiology
MOLECULAR BIOLOGY
PCR
Physics, chemistry, biochemistry and biology of agricultural and forest soils
PRESERVACION
PRESERVATION
SOIL
Soil science
SOL
SUELO
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Summary:The aim of this study was to select a method for preserving bacterial DNA in soil samples in cases where there are no possibilities of using freezing or cooling methods. To overcome this difficulty, we hypothesized that adding absolute ethanol to soil samples could be as successful to preserve bacteria as it is to preserve insect or tissue samples for molecular studies. In an attempt to test this assumption, we compared four conservation conditions. After inoculation of soil samples with Escherichia coli, they were either kept at 28 °C, stored in the cold (4 °C), dried at 60 °C, or treated with absolute ethanol. The relative effectiveness of the methods was evaluated by using both DNA recoveries and bacterial 16S rDNA amplification as criteria. Two kinds of soils, i.e. sandy clay and clayey soil, were used. Soil conservation was tested for seven time periods ranging from 2 d to 1 year after bacterial inoculation. Results showed that cold conservation or addition of absolute ethanol to the samples yielded similar DNA recoveries. For these treatments, successful amplifications are still obtained after one year of conservation. The ethanol conservation of soil samples appears to be the easiest method to preserve the bacterial DNA because of its reliability and field convenience.
ISSN:1164-5563
DOI:10.1016/S1164-5563(00)00044-3