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Technical considerations for the use of N-15-DNA stable-isotope probing for functional microbial activity in soils
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UNSPECIFIED (2005) Technical considerations for the use of N-15-DNA stable-isotope probing for functional microbial activity in soils. In: Joint Meeting of the European-Stabel-Isotope-Users-Group, Vienna, AUSTRIA, AUG 30-SEP 03, 2004. Published in: RAPID COMMUNICATIONS IN MASS SPECTROMETRY, 19 (11). pp. 1424-1428. doi:10.1002/rcm.1908 ISSN 0951-4198.
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Official URL: http://dx.doi.org/10.1002/rcm.1908
Abstract
Stable-isotope DNA probing is a culture-independent technique that may provide a link between function and phylogeny of active microorganisms. The technique has been used in association with C-13 substrates while here we evaluate feasibility and limitations of N-15-DNA stable-isotope probing (SIP) using labelled and unlabelled pure microbial cultures or soil extracts. Our results showed that N-15-DNA probing is feasible for cultures as well as soil samples. Limitations of N-15-DNA-SIP are (a) the need for relatively large quantities of DNA to visualise bands (although molecular resolution is much higher) and (b) N-15-DNA enrichment needed to ideally be > 50 at%; however, this requirement can be lowered to approx. 40 atom% N-15 with pure cultures using a modified CsCl centrifugation method (140Kg for 69h). These advances in N-15-DNA-SIP methodology open new opportunities to trace active microbial populations utilising specific N substrates in situ. Copyright © 2005 John Wiley & Sons, Ltd.
Item Type: | Conference Item (UNSPECIFIED) | ||||
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Subjects: | Q Science > QD Chemistry Q Science > QC Physics |
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Journal or Publication Title: | RAPID COMMUNICATIONS IN MASS SPECTROMETRY | ||||
Publisher: | JOHN WILEY & SONS LTD | ||||
ISSN: | 0951-4198 | ||||
Official Date: | 2005 | ||||
Dates: |
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Volume: | 19 | ||||
Number: | 11 | ||||
Number of Pages: | 5 | ||||
Page Range: | pp. 1424-1428 | ||||
DOI: | 10.1002/rcm.1908 | ||||
Publication Status: | Published | ||||
Title of Event: | Joint Meeting of the European-Stabel-Isotope-Users-Group | ||||
Location of Event: | Vienna, AUSTRIA | ||||
Date(s) of Event: | AUG 30-SEP 03, 2004 |
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