Abstract：
:Soil is a complex environment considered as one of the main reservoirs of microbial diversity. However, the inability to cultivate most soil bacteria hampered fundamental attempts to determine the diversity of the prokaryotic world and limited its industrial exploitation. In the last 20 years, new methods have been developed to overcome these limitations based on the direct extraction of DNA from bacteria in their natural environment. In addition to fundamental research, the cloning of the extracted DNA for the development of metagenomic DNA clone libraries offers possibilities to discover novel bio-molecules through the expression of genes from uncultivated bacteria in surrogate bacterial hosts. However, such objectives require adapting DNA extraction methods and cloning strategies in order that entire gene clusters encoding biosynthetic pathway for secondary metabolites can be cloned. In this paper, we report that the size of DNA fragments extracted from soil varied in a range between less than 100 kb and more than 400 kb depending on the soil. The relatively limited size of DNA fragments extracted from some soil was not only due to mechanical, chemical or enzymatic shearing of the DNA during the extraction process but partly to the microbial growth status. Stimulating bacteria in situ by providing nutrients to the soil improved the size of extracted DNA, but it modified the bacterial community structure.
journal_name
J Microbiol Methodsjournal_title
Journal of microbiological methodsauthors
Bertrand H,Poly F,Van VT,Lombard N,Nalin R,Vogel TM,Simonet Pdoi
10.1016/j.mimet.2005.01.003subject
Has Abstractpub_date
2005-07-01 00:00:00pages
1-11issue
1eissn
0167-7012issn
1872-8359pii
S0167-7012(05)00022-9journal_volume
62pub_type
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