Soil conservation for DNA preservation for bacterial molecular studies

被引:26
作者
Harry, M [1 ]
Gambier, B [1 ]
Garnier-Sillam, E [1 ]
机构
[1] Univ Paris 12, UFR Sci, Lab Biol Sols & Eaux, F-94000 Creteil, France
关键词
soil; conservation; DNA recovery; bacteria; PCR;
D O I
10.1016/S1164-5563(00)00044-3
中图分类号
Q14 [生态学(生物生态学)];
学科分类号
071012 ; 0713 ;
摘要
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. Ln 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 degrees C, stored in the cold (4 degrees C), dried at 60 degrees 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. (C) 2000 Editions scientifiques et medicales Elsevier SAS.
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页码:51 / 55
页数:5
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