Efficient recovery of environmental DNA for expression cloning by indirect extraction methods

Using direct and cell extraction-based (indirect) isolation methods, DNA was obtained from environmental samples with largely differing characteristics (loam soil, sand soil, sediment, activated sludge, and compost) and evaluated with respect to the comprised bacterial diversity and its suitability...

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Bibliographic Details
Published in:FEMS microbiology ecology 2003-05, Vol.44 (2), p.153-163
Main Authors: Gabor, Esther M, de Vries, Erik J, Janssen, Dick B
Format: Article
Language:English
Subjects:
Activated sludge
Bacteria
Cloning
Construction methods
Deoxyribonucleic acid
DNA
E coli
Ecology
Electrophoresis
Environmental DNA
Gel electrophoresis
Gene banks
Gene expression
Inserts
Loam soils
Lysis
Microbiology
Soils
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Summary:Using direct and cell extraction-based (indirect) isolation methods, DNA was obtained from environmental samples with largely differing characteristics (loam soil, sand soil, sediment, activated sludge, and compost) and evaluated with respect to the comprised bacterial diversity and its suitability for expression cloning in Escherichia coli. Indirect DNA extraction methods yielded 10 to 100-fold lower amounts of DNA than direct procedures, but the bacterial diversity of DNA recovered by indirect means was distinctly higher as shown by denaturing gradient gel electrophoresis. Furthermore, much lower amounts of eukaryotic DNA were co-extracted if cell extraction-based methods were used (
ISSN:0168-6496
1574-6941
DOI:10.1016/S0168-64960200462-2