Figure 5.

Biophysical and ¹H NMR characterization of 19wt and its loop mutants. (A) ¹H NMR spectra at 700 MHz and 298 K. The oligonucleotides were prepared at 120–200 μM in KPi buffer and 10% D₂O. (B) Non-denaturing 15% PAGE. Samples were prepared at 70 μM in 5K buffer. (C) CD spectra. All mutants with G5A or G5T mutations (M1–M3 and M5–M7) have a slightly increased CD intensity at 295 nm while the CD signatures of M4 and M8 are nearly identical to that of 19wt. (D) TDS. (E) Bar graph of ΔT_m (relative to 19wt) determined via CD melting studies. For C–E, the samples were 2.5–4.5 μM in 5K buffer.