Figure 4

Melting curves after change of buffer and effect on Tm of probe concentrations. (A) Fluorescence of ATTO495-ATTO647N antiparallel duplex (AD) FRET at 670 nm from pH 5.5 to 8.0 with 0.5 μM of each probe in sodium phosphate buffer. (B) Fluorescence of ATTO495-ATTO647N parallel triplex (PT) FRET at 670 nm from pH 4.5 to 6.0 with 1.0 μM of each probe in sodium acetate buffer. (C) ATTO495-ATTO647N antiparallel duplex FRET at 670 nm in sodium phosphate buffer pH 7.0 and probe concentrations from 0.125 μM to 1.00 μM of each probe. (D) ATTO495-ATTO647N parallel triplex FRET at 670 nm in sodium acetate buffer pH 5.0 and probe concentrations from 0.125 μM to 1.50 μM of each probe.