Characterization of the dsMango variants. (A–C) Fluorescence saturation curves with 25 nM TO1-b and variable RNA concentrations. Shaded regions show 68% CI of the curve fit. Constructs are grouped based on whether the first stem is at (A) the i-iv corner, (B) the ii-i corner, or (C) the iii-ii corner. (D) Relative maximum fluorescence (Fmax) of the double-stemmed variants. (E) Fraction of dsMango i-iv ii-i in free solution over a range of TO1-b concentrations. Curve fit line represents least squares fit using Equation 1 (see Materials and Methods). KD derived from curve fitting is indicated by the dotted line. The shaded region representing the 95% CI is determined from error curve analysis.
