Site-directed mutagenesis of RfaHCC

Plasmid pET19mod_rfaH-F51C-S139C encoding RfaH^CC was generated by successive site-directed mutagenesis according to the QuickChange Site-Directed Mutagenesis Kit protocol, using pET19mod_rfaH as a template^{33 (link)}. All other plasmids were constructed by standard molecular biology approaches with restriction and modification enzymes from New England Biolabs. Sequences of all plasmids were confirmed by Sanger sequencing either at the Genomics Shared Resource Facility, Ohio State University, USA or at Eurofins Genomics, Germany.

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Zuber P.K., Said N., Hilal T., Wang B., Loll B., González-Higueras J., Ramírez-Sarmiento C.A., Belogurov G.A., Artsimovitch I., Wahl M.C, & Knauer S.H. (2024). Concerted transformation of a hyper-paused transcription complex and its reinforcing protein. Nature Communications, 15, 3040.

Publication 2024

restriction and modification enzymes

Corresponding Organization :

Other organizations : University of Bayreuth, Freie Universität Berlin, The Ohio State University, Pontificia Universidad Católica de Chile, University of Turku

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Variable analysis

independent variables

Site-directed mutagenesis to generate plasmid pET19mod_rfaH-F51C-S139C encoding RfaHCC

dependent variables

Not explicitly mentioned

control variables

PET19mod_rfaH as a template for site-directed mutagenesis

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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