NDLI: A Comparison of the Use of In Vitro–Transcribed and Native rRNA for the Quantification of Microorganisms in the Environment

Content Provider	Springer Nature Link
Author	McMahon, K.D. Stahl, D.A. Raskin, L.
Abstract	Nearly full-length, small subunit (SSU) rRNA was transcribed in vitro from clones of SSU rDNA genes. Comparing the use of in vitro–transcribed and native rRNA indicated that, when in vitro–transcribed rRNA was used as a standard for quantitative hybridizations with oligonucleotide probes, the population was consistently underestimated. The population abundance was expressed as a percentage of specific target SSU rRNA (determined with a specific oligonucleotide probe), relative to the total SSU rRNA (measured with a universal probe). Differences in hybridization signals could be related to specific probe target locations and rRNA denaturation conditions, suggesting that higher order structure is important in quantitative membrane hybridizations. Therefore, in vitro–transcribed rRNA cannot always be used for the absolute quantification of microbial populations, but can be employed as a standard to quantify shifts in population abundance over time, and to compare community structure in various environments.
Starting Page	362
Ending Page	371
Page Count	10
File Format	PDF
ISSN	00953628
Journal	Microbial Ecology
Volume Number	36
Issue Number	3-4
e-ISSN	1432184X
Language	English
Publisher	Springer-Verlag
Publisher Date	1998-11-01
Publisher Place	Berlin, Heidelberg
Access Restriction	Subscribed
Content Type	Text
Resource Type	Article
Subject	Soil Science Ecology Ecology, Evolution, Behavior and Systematics

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A Comparison of the Use of In Vitro–Transcribed and Native rRNA for the Quantification of Microorganisms in the Environment