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Updated: Jun 3, 2026

In Situ Hybridization for the Precise Localization of Transcripts in Plants
Published on: November 23, 2011
In vitro transcription in plant nuclei
1Division of Biology and Environmental Sciences, The Hatfield Polytechnic, Hertfordshire, UK.
Abstract:
Isolated plant nuclei can be used for fundamental studies on the transcription apparatus. Total RNA polymerase activity can be measured using plant nuclei, and by using different α-amanitin concentrations in the enzyme assay, the individual RNA polymerase I, II, and in activities can be measured (1-5). The assay procedure involves the incubation of nuclei in the presence of the four substrates for RNA synthesis: ATP, GTP, CTP, and UTP. If one of these precursors is supplied as a radio-labeled molecule, transcription can be detected as incorporation of radioactivity into acid-insoluble material. Following incubation, transcription products are precipitated with TCA, collected and washed on glass fiber filter discs, and counted by liquid scintillation counting.
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