title: In vitro transcription by RNA polymerase II in extracts of Xenopus oocytes, eggs, and somatic cells.
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.
authors.affiliation: Laboratory of Molecular Embryology, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892.
authors.forename: T
authors.initials: T
authors.lastname: Toyoda
authors.forename: A P
authors.initials: AP
authors.lastname: Wolffe
mesh_terms: D000818:Animals; D001483:Base Sequence; D002457:Cell Extracts; D002478:Cells, Cultured; D008969:Molecular Sequence Data; D009865:Oocytes / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic; D012319:RNA Polymerase II / Q000378:metabolism*; D013696:Temperature; D014158:Transcription, Genetic; D014982:Xenopus laevis
chemical_list: D002457:Cell Extracts; D012319:RNA Polymerase II