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Protocol for monitoring and analyzing single nucleotide incorporation by S. cerevisiae RNA polymerases.
Jacobs, Ruth Q; Bellis, Nathan F; Lucius, Aaron L; Schneider, David A.
Afiliación
  • Jacobs RQ; Deparment of Biochemistry and Molecular Genetics, School of Medicine, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
  • Bellis NF; Deparment of Biochemistry and Molecular Genetics, School of Medicine, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
  • Lucius AL; Department of Chemistry, University of Alabama at Birmingham, Birmingham, AL 35294, USA. Electronic address: allucius@uab.edu.
  • Schneider DA; Deparment of Biochemistry and Molecular Genetics, School of Medicine, University of Alabama at Birmingham, Birmingham, AL 35294, USA. Electronic address: dschneid@uab.edu.
STAR Protoc ; 4(2): 102191, 2023 Mar 24.
Article en En | MEDLINE | ID: mdl-36964908
Here we present an optimized protocol for monitoring and analyzing single nucleotide incorporation by RNA polymerases. This protocol describes the assembly of Saccharomyces cerevisiae RNA polymerase I elongation complexes in a promoter-independent system in vitro. We describe how to collect a time course using a quench-flow, a rapid mixing instrument, and subsequently resolve reactions on a polyacrylamide gel. Finally, we detail how to quantify the gel images. For complete details on the use and execution of this protocol, please refer to Appling et al. (2015).1.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: STAR Protoc Año: 2023 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: STAR Protoc Año: 2023 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos