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PMID: 12438682 Published · ppublish English Comparative Study Journal Article

Cultivating the uncultured.

Zengler K, Toledo G, Rappe M, Elkins J, Mathur EJ, Short JM, Keller M

Abstract

The recent application of molecular phylogeny to environmental samples has resulted in the discovery of an abundance of unique and previously unrecognized microorganisms. The vast majority of this microbial diversity has proved refractory to cultivation. Here, we describe a universal method that provides access to this immense reservoir of untapped microbial diversity. This technique combines encapsulation of cells in gel microdroplets for massively parallel microbial cultivation under low nutrient flux conditions, followed by flow cytometry to detect microdroplets containing microcolonies. The ability to grow and study previously uncultured organisms in pure culture will enhance our understanding of microbial physiology and metabolic adaptation and will provide new sources of microbial metabolites. We show that this technology can be applied to samples from several different environments, including seawater and soil.

MeSH Terms
Bacteria/classification,genetics,isolation & purification Bacteriological Techniques Drug Compounding Flow Cytometry Fresh Water/microbiology Gels Ghana Kenya Particle Size Phylogeny Polymerase Chain Reaction RNA, Ribosomal/genetics RNA, Ribosomal, 16S/genetics Seawater/microbiology Soil Microbiology Species Specificity Water Microbiology
Chemicals
Gels RNA, Ribosomal RNA, Ribosomal, 16S
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Zengler Karsten
Diversa Corporation, San Diego, CA 92121, USA.
Toledo Gerardo
Rappe Michael
Elkins James
Mathur Eric J
Short Jay M
Keller Martin
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
Published
2002-11-26
Epub
2002-00-18
Pages
15681-6
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC137776
Subset
IM
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