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In vitro termination assay with c-di-GMP. (A) An in vitro transcription reaction with E. coli RNA polymerase and sigma 70 was performed on a DNA template containing the sequence upstream of tfoY and part of the tfoY open reading frame. Mutations to the riboswitch are indicated. (B) A similar in vitro transcription reaction was performed with a class II riboswitch from Clostridium difficile. c-di-GMP was added at a concentration of 0.1 mM.
