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Nat Protoc. 2008;3(8):1261-9. doi: 10.1038/nprot.2008.102.

Cultivating previously uncultured soil bacteria using a soil substrate membrane system.

Author information

  • 1School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, New South Wales 2052, Australia. b.ferrari@unsw.edu.au

Abstract

Most bacteria are recalcitrant to traditional cultivation in the laboratory. The soil substrate membrane system provides a simulated environment for the cultivation of previously undescribed soil bacteria as microcolonies. The system uses a polycarbonate membrane as a solid support for growth and soil extract as the substrate. Diverse microcolonies can be visualized using total bacterial staining combined with fluorescence in situ hybridization (FISH) after 7-10-d incubation. Molecular typing shows that the majority of microcolony-forming bacteria recovered using this protocol were resistant to growth using standard methods. The protocol takes <4 h of bench time over the 10-d period.

PMID:
18714294
DOI:
10.1038/nprot.2008.102
[PubMed - indexed for MEDLINE]
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