University of Hertfordshire

From the same journal

In vitro transcription in plant nuclei

Research output: Contribution to journalArticle

  • R. D. Henfrey
  • R. J. Slater
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Original languageEnglish
Number of pages11
Pages (from-to)453-63
JournalMethods in Molecular Biology
Journal publication date1988
Volume4
DOIs
Publication statusPublished - 1988

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.

ID: 1576865