Table 3. Gibbs free energy (kcal/mol) for base-pair stability of the WT and mutant pri-miR156a constructs at 23°C.

Base pair		WT	A9C	A9AC	A10G	A10CG
C15·G98	ΔG°bp	–6.87 ± 0.14	–7.44 ± 0.11	–6.86 ± 0.07	–6.83 ± 0.12	–7.43 ± 0.24
	ΔΔG°bp	–	–0.57	0.01	0.04	–0.56
	ΔΔG‡op	–	–0.37	0.05	0.26	–0.04
	ΔΔG‡cl	–	–0.94	0.06	0.30	–0.60
U16·A97	ΔG°bp	–6.35 ± 0.05	–6.98 ± 0.04	–6.39 ± 0.03	–6.48 ± 0.08	–7.07 ± 0.05
	ΔΔG°bp	–	–0.63	–0.04	–0.13	–0.72
	ΔΔG‡op	–	–0.96	–0.08	–0.07	–0.66
	ΔΔG‡cl	–	–1.59	–0.12	–0.20	–1.38
U16·A97 + C15·G98	ΔG°bp	–13.22	–14.42	–13.25	–13.31	–14.50
	ΔΔG°bp	–	–1.20	–0.03	–0.09	–1.28
	ΔΔG‡op	–	–1.33	–0.03	0.19	–0.70
	ΔΔG‡cl	–	–2.53	–0.06	0.10	–1.98
G17·C96	ΔG°bp	–8.44 ± 0.43	–8.67 ± 0.32	–8.38 ± 0.22	–8.61 ± 0.48	–8.57 ± 0.39
	ΔΔG°bp	–	–0.23	0.06	–0.17	–0.13
	ΔΔG‡op	–	0.39	–0.06	0.49	0.47
	ΔΔG‡cl	–	0.16	0.00	0.32	0.34
A18·U95	ΔG°bp	–7.05 ± 0.18	–7.45 ± 0.11	–7.01 ± 0.07	–7.57 ± 0.50	–7.37 ± 0.13
	ΔΔG°bp	–	–0.40	0.04	–0.52	–0.32
	ΔΔG‡op	–	–0.10	0.08	–0.15	0.09
	ΔΔG‡cl	–	–0.50	0.12	–0.67	–0.23
C19·G94	ΔG°bp	–7.56 ± 0.15	–8.00 ± 0.19	–7.74 ± 0.18	–7.94 ± 0.43	–7.73 ± 0.20
	ΔΔG°bp	–	–0.44	–0.18	–0.38	–0.17
	ΔΔG‡op	–	0.07	–0.11	–0.04	–0.17
	ΔΔG‡cl	–	–0.37	–0.29	–0.42	–0.34

^aΔG°_bp = –ΔG°_opening = RTln(K_op), T = 296.15 K.

^bΔΔG°_bp = ΔG°_bp,mut – ΔG°_bp,WT.

^cΔΔG^‡_op = ΔG^‡_op,mut – ΔG^‡_op,WT = –RTln(k_op,mut/k_op,WT) = RTln(τ_0,mut/τ_0,WT), T = 296.15 K.

^dΔΔG^‡_cl = ΔG^‡_cl,mut – ΔG^‡_cl,WT = –RTln(k_cl,mut/k_cl,WT) = RTln(τ_open,mut/τ_open,WT), T = 296.15 K.