3.2. UV Melting Studies

The single-stranded oligonucleotide concentrations were calculated based on their absorbance at 85 °C, and the extinction coefficients were calculated using the OligoAnalyzer tool (Integrated DNA Technologies). UV melting analysis was performed for nine different concentrations of each oligonucleotide in the range of 10⁻⁴ to 10⁻⁶ M. The oligonucleotides were dissolved in buffer containing 100 mM potassium chloride (KCl), 20 mM sodium cacodylate and 0.5 mM Na₂EDTA (pH 7.0). The buffer was degassed at an elevated temperature before the measurements. Absorbance versus temperature curves were obtained using the UV melting method at 295 nm with the temperature range of 95 to 3 °C and a temperature decrease of 0.2 °C/min (Supplementary Data, Figure S9) using a JASCO V-650 (Cremella (LC) Italy) spectrophotometer equipped with a thermoprogrammer. The thermodynamic parameters were analyzed and determined using MeltWin 3.5 software. The melting temperatures calculated for the 10⁻⁴ M concentration of the oligonucleotide are denoted by T_M, and the melting points for any other concentration of oligonucleotide are denoted by T_m.