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  • Title: Cultivating previously uncultured soil bacteria using a soil substrate membrane system.
    Author: Ferrari BC, Winsley T, Gillings M, Binnerup S.
    Journal: Nat Protoc; 2008; 3(8):1261-9. PubMed ID: 18714294.
    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.
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