title: Direct competition assay for transcription fidelity.
abstract: Accurate transcription is essential for faithful information flow from DNA to RNA and to the protein. Mechanisms of cognate substrate selection by RNA polymerases are currently elucidated by structural, genetic, and biochemical approaches. Here, we describe a fast and reliable approach to quantitative analyses of transcription fidelity, applicable to analyses of RNA polymerase selectivity against misincorporation, incorporation of dNMPs, and chemically modified rNMP analogues. The method is based on different electrophoretic mobility of RNA oligomers of the same length but differing in sequence.
authors: NCI Center for Cancer Research, Frederick, MD, 21702, USA.
authors: Lucyna
authors: L
authors: Lubkowska
authors: Maria L
authors: ML
authors: Kireeva
mesh_terms: D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay; D008967:Molecular Biology / Q000379:methods*; D046912:Multiprotein Complexes / Q000378:metabolism*; D013379:Substrate Specificity; D014158:Transcription, Genetic / Q000235:genetics / Q000502:physiology*; D035602:Transcriptional Elongation Factors / Q000378:metabolism*
chemical_list: D046912:Multiprotein Complexes; D035602:Transcriptional Elongation Factors; D012321:DNA-Directed RNA Polymerases