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2.3 Physical measurements
1H NMR spectra were recorded on an Avance-400 spectrometer with d6-DMSO (dimethyl sulfoxide) as the solvent at room temperature and TMS (tetramethylsilane) as the internal standard. UV–Vis (UV–visible) spectra were recorded on a Perkin–Elmer Lambda-25 spectrophotometer, and emission spectra were recorded on a PerkinElmer LS-55 luminescence spectrometer at room temperature. Electrospray mass spectra (ES-MS) were recorded on a LQC system (Finngan MAT, USA) using CH3CN as the mobile phase.
The absorption titration of ruthenium(II) complexes in buffer was carried out by adding the DNA stock solution to a fixed concentration of ruthenium. Ruthenium solutions employed were 10 μM in concentration and calf thymus DNA was added to a ratio of 1.27:1 [DNA]/[Ru]. Ruthenium-DNA solutions were allowed to incubate for 5 min before the absorption spectra were recorded. The intrinsic binding constants Kb of Ru (II) complexes to DNA were calculated from equation [14]:
[DNA] / (εa -f) = [DNA] / (b -f) + 1 / [Kb(b -f)]
Where [DNA] is the concentration of DNA in base pairs, εa, εf and εb correspond to the apparent absorption coefficient Aobsd/[Ru], the extinction coefficient for the ruthenium complex in the fully bound form, respectively. In plots of [DNA]/( εa -εf ) versus [DNA], Kb is given by the ratio of slope to the intercept.
Viscosity measurements were carried out by using an Ubbelodhe viscometer maintained a constant temperature at 28.0 (±0.1) C in thermostatic water-bath. CT-DNA samples approximately 200 base pairs in average length were prepared by sonicating in order to minimize complexities arising from DNA flexibility [15]. Flow time was measured with a digital stopwatch, and each sample was measured three times, and an average flow time was calculated. Data were presented as (/0) 1/3 versus binding ratio [16], where is the viscosity of CT-DNA in the presence of complex, and 0 is the viscosity of CT-DNA alone.
2.3 Physical measurements
1H NMR spectra were recorded on an Avance-400 spectrometer with d6-DMSO (dimethyl sulfoxide) as the solvent at room temperature and TMS (tetramethylsilane) as the internal standard. UV–Vis (UV–visible) spectra were recorded on a Perkin–Elmer Lambda-25 spectrophotometer, and emission spectra were recorded on a PerkinElmer LS-55 luminescence spectrometer at room temperature. Electrospray mass spectra (ES-MS) were recorded on a LQC system (Finngan MAT, USA) using CH3CN as the mobile phase.
The absorption titration of ruthenium(II) complexes in buffer was carried out by adding the DNA stock solution to a fixed concentration of ruthenium. Ruthenium solutions employed were 10 μM in concentration and calf thymus DNA was added to a ratio of 1.27:1 [DNA]/[Ru]. Ruthenium-DNA solutions were allowed to incubate for 5 min before the absorption spectra were recorded. The intrinsic binding constants Kb of Ru (II) complexes to DNA were calculated from equation [14]:
[DNA] / (εa -f) = [DNA] / (b -f) + 1 / [Kb(b -f)]
Where [DNA] is the concentration of DNA in base pairs, εa, εf and εb correspond to the apparent absorption coefficient Aobsd/[Ru], the extinction coefficient for the ruthenium complex in the fully bound form, respectively. In plots of [DNA]/( εa -εf ) versus [DNA], Kb is given by the ratio of slope to the intercept.
Viscosity measurements were carried out by using an Ubbelodhe viscometer maintained a constant temperature at 28.0 (±0.1) C in thermostatic water-bath. CT-DNA samples approximately 200 base pairs in average length were prepared by sonicating in order to minimize complexities arising from DNA flexibility [15]. Flow time was measured with a digital stopwatch, and each sample was measured three times, and an average flow time was calculated. Data were presented as (/0) 1/3 versus binding ratio [16], where is the viscosity of CT-DNA in the presence of complex, and 0 is the viscosity of CT-DNA alone.
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