Fig. 2.

Sequence mismatches and torque affect the stability of R-loop intermediates. (A) (Left) Example trace showing R-loop formation with λ1 sgRNA. Blue, raw 5-kHz data; yellow, change-point analysis (43, 44) fit. (Right) Histogram of change in equilibrium twist ($\mathrm{\Delta }{\theta }_0$, in base pairs) as a function of imposed twist. Data from multiple cycles using measurement protocols as in Fig. 1E are block averaged in 20-ms bins and aligned in twist; plots contain data from multiple experiment repetitions and duplicate sgRNA preparations (Materials and Methods). (B–F) Data as in A for 13-20mm, 18CA, 1-4mm, 3CA, and 8TA sgRNAs. Mismatched bases are highlighted in red. Cartoon in B illustrates the orientation of the DNA and sgRNA segments.