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  • Title: In vitro transcription by RNA polymerase II in extracts of Xenopus oocytes, eggs, and somatic cells.
    Author: Toyoda T, Wolffe AP.
    Journal: Anal Biochem; 1992 Jun; 203(2):340-7. PubMed ID: 1416031.
    Abstract:
    We describe procedures for preparing extracts of Xenopus oocytes, eggs, and somatic cells that will accurately transcribe class II genes. A variety of viral and Xenopus promoters direct the accurate initiation of transcription by RNA polymerase II in these extracts. Optimal ionic conditions (100-200 mM KCl, 12 mM MgCl2), template concentration (20-40 micrograms/ml), incubation time (30-60 min), and temperature (25 degrees C) for class II gene transcription are described.
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