Run-on transcription as a method for the analysis of mitochondrial genome expression

Run-on transcription as a method for the analysis of mitochondrial genome expression The main stages of the run-on transcription method as applied to the quantification of mitochondrial gene transcription are described. As distinct from widely applied methods of northern hybridization and PCR after reverse transcription, this method permits studying the effects of exogenous and endogenous factors on the rate of mitochondrial gene transcription. The main method stages are (1) isolation of intact organelles; (2) preparation of nylon membranes with gene-specific fragments of DNA; (3) transcription in the lysates of these organelles in the presence of 32P-labeled RNA precursors; and (4) hybridization of newly synthesized labeled transcripts with DNA fragments of tested mitochondrial genes loaded on the nylon membrane. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Russian Journal of Plant Physiology Springer Journals

Run-on transcription as a method for the analysis of mitochondrial genome expression

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Publisher
SP MAIK Nauka/Interperiodica
Copyright
Copyright © 2011 by Pleiades Publishing, Ltd.
Subject
Life Sciences; Plant Sciences ; Plant Physiology
ISSN
1021-4437
eISSN
1608-3407
D.O.I.
10.1134/S1021443711010031
Publisher site
See Article on Publisher Site

Abstract

The main stages of the run-on transcription method as applied to the quantification of mitochondrial gene transcription are described. As distinct from widely applied methods of northern hybridization and PCR after reverse transcription, this method permits studying the effects of exogenous and endogenous factors on the rate of mitochondrial gene transcription. The main method stages are (1) isolation of intact organelles; (2) preparation of nylon membranes with gene-specific fragments of DNA; (3) transcription in the lysates of these organelles in the presence of 32P-labeled RNA precursors; and (4) hybridization of newly synthesized labeled transcripts with DNA fragments of tested mitochondrial genes loaded on the nylon membrane.

Journal

Russian Journal of Plant PhysiologySpringer Journals

Published: Jan 8, 2011

References

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