A simple and rapid method to detect plant siRNAs using nonradioactive probes

A simple and rapid method to detect plant siRNAs using nonradioactive probes Small interfering RNAs (siRNAs) are key molecules in RNA silencing, which includes posttranscriptional gene silencing, cosuppression, quelling, and RNA interference. The presence of siRNAs indicates RNA silencing in cells. We present a method of detecting siRNAs using nonradioactive probes that involves isolating the small RNA fraction, separating siRNAs using denaturing gel electrophoresis, and performing a Northern blot analysis under low-stringency hybridization conditions. We used digoxigenin-labeled DNA probes for hybridization and detected siRNAs in petunia and rice plants exhibiting silenced phenotypes. This method is a simple and rapid way to detect siRNAs without using radioisotopes. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Plant Molecular Biology Reporter Springer Journals

A simple and rapid method to detect plant siRNAs using nonradioactive probes

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Publisher
Springer Journals
Copyright
Copyright © 2003 by International Society for Plant Molecular Biology
Subject
Life Sciences; Plant Sciences; Plant Breeding/Biotechnology; Proteomics; Metabolomics; Bioinformatics
ISSN
0735-9640
eISSN
1572-9818
D.O.I.
10.1007/BF02773396
Publisher site
See Article on Publisher Site

Abstract

Small interfering RNAs (siRNAs) are key molecules in RNA silencing, which includes posttranscriptional gene silencing, cosuppression, quelling, and RNA interference. The presence of siRNAs indicates RNA silencing in cells. We present a method of detecting siRNAs using nonradioactive probes that involves isolating the small RNA fraction, separating siRNAs using denaturing gel electrophoresis, and performing a Northern blot analysis under low-stringency hybridization conditions. We used digoxigenin-labeled DNA probes for hybridization and detected siRNAs in petunia and rice plants exhibiting silenced phenotypes. This method is a simple and rapid way to detect siRNAs without using radioisotopes.

Journal

Plant Molecular Biology ReporterSpringer Journals

Published: Sep 3, 2012

References

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