Methodological considerations for the use of stable isotope probing in microbial ecology
Methodological considerations for the use of stable isotope probing in microbial ecology
Stable isotope probing (SIP) is a method used for labeling uncultivated microorganisms in environmental samples or directly in field studies using substrate enriched with stable isotope (e.g., 13C). After consumption of the substrate, the cells of microorganisms that consumed the substrate become enriched in the isotope. Labeled biomarkers, such as phospholipid-derived fatty acid (PLFA), ribosomal RNA, and DNA can be analyzed with a range of molecular and analytical techniques, and used to identify and characterize the organisms that incorporated the substrate. The advantages and disadvantages of PLFA-SIP, RNA-SIP, and DNA-SIP are presented. Using examples from our laboratory and from the literature, we discuss important methodological considerations for a successful SIP experiment.
435–442
Neufeld, Josh D
97a99cce-a614-441b-ab85-dbae37e3c4ba
Dumont, Marc
afd9f08f-bdbb-4cee-b792-1a7f000ee511
Vohra, Jyotsna
d5c7571e-5ec2-46d6-a77c-64e25a7f2413
Murrell, J. Colin
244a92ff-dbe1-41cf-9e65-baacbc4a90cf
28 October 2006
Neufeld, Josh D
97a99cce-a614-441b-ab85-dbae37e3c4ba
Dumont, Marc
afd9f08f-bdbb-4cee-b792-1a7f000ee511
Vohra, Jyotsna
d5c7571e-5ec2-46d6-a77c-64e25a7f2413
Murrell, J. Colin
244a92ff-dbe1-41cf-9e65-baacbc4a90cf
Neufeld, Josh D, Dumont, Marc, Vohra, Jyotsna and Murrell, J. Colin
(2006)
Methodological considerations for the use of stable isotope probing in microbial ecology.
Microbial Ecology, 53, .
(doi:10.1007/s00248-006-9125-x).
Abstract
Stable isotope probing (SIP) is a method used for labeling uncultivated microorganisms in environmental samples or directly in field studies using substrate enriched with stable isotope (e.g., 13C). After consumption of the substrate, the cells of microorganisms that consumed the substrate become enriched in the isotope. Labeled biomarkers, such as phospholipid-derived fatty acid (PLFA), ribosomal RNA, and DNA can be analyzed with a range of molecular and analytical techniques, and used to identify and characterize the organisms that incorporated the substrate. The advantages and disadvantages of PLFA-SIP, RNA-SIP, and DNA-SIP are presented. Using examples from our laboratory and from the literature, we discuss important methodological considerations for a successful SIP experiment.
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Accepted/In Press date: 12 June 2006
Published date: 28 October 2006
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Local EPrints ID: 480333
URI: http://eprints.soton.ac.uk/id/eprint/480333
ISSN: 0095-3628
PURE UUID: 218e2114-76da-4bb4-9cd5-d7a38f6a5a6c
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Date deposited: 01 Aug 2023 17:23
Last modified: 18 Mar 2024 03:33
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Author:
Josh D Neufeld
Author:
Jyotsna Vohra
Author:
J. Colin Murrell
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