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Transcription assay using plasmids pE3SP-shLuc and pU6-shLuc in nuclear extracts

figure

posted on 2011-12-31, 03:44 authored by M. S. Brenz Verca, Peter Weber, Christine Mayer, Cornelia Graf, Damián Refojo, Ralf Kühn, Ingrid Grummt, Beat Lutz

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Taken from "Development of a species-specific RNA polymerase I-based shRNA expression vector"

Nucleic Acids Research 2006;35(2):e10-e10.

Published online 07 Dec 2006

PMCID:PMC1802596.

Assays were done in absence or presence of α-amanitin, a specific inhibitor of Pol II- and Pol III-dependent transcription. () The expected band of 60 nt, corresponding to the expressed shRNA. () Additional fragment of 150 nt in length, resulting from pU6-shLuc (Pol III-dependent) transcription. () 5S rRNA transcript (Pol III-dependent), used as a positive control for assay, and inhibition by α-amanitin. Asterisk indicates an unspecific signal, inherent to the transcription from nuclear extract of FM3A cells.