NMR spectra were obtained with Agilent Technologies VNMRS 800 MHz and DD2 600 MHz NMR spectrometers equipped with triple resonance cold probe. Standard 1D 1H-NMR spectra were acquired with the use of DPFGSE or watergate 3919 solvent suppression. Diffusion coefficient measurements were performed by a spin-echo pulse sequence with PFG gradient strengths between 1.3 and 32.5 Gcm−1. NOESY spectra were recorded using mixing times of 100, 80, 40 and 20 ms. TOCSY spectrum was recorded using mixing times of 80 ms. 1H-NMR spectra were acquired at 0, 5, 7, 15, 20, 25, 37 and 40 °C. Effect of different pHs was observed by recording 1H-NMR spectra at pH 4.5, 5.0, 5.5, 6.0, 6.5 and 7.0. Effect of molecular crowding conditions was observed by recording 1H-NMR spectra in the presence of 10% w/v PEG (8000 MW) in autoclaved H2O. In experiments with K+ ions 3M KCl was diluted into the sample up to the desired concentration of K+ ions. 1D and 2D NMR spectra were processed and analysed using VNMRJ (Varian Inc.), MestReNova and Sparky (UCSF) software. All NMR spectra were acquired on 600 MHz NMR spectrometer unless stated otherwise. DSS (4,4-dimethyl-4-silapentane-1-sulfonic acid) was used as external reference in all NMR spectra.
Vnmrs 800 mhz
Manufactured by Agilent Technologies
The VNMRS 800 MHz is a high-resolution nuclear magnetic resonance (NMR) spectrometer designed for advanced analytical and research applications. It features an 800 MHz superconducting magnet and provides exceptional performance and sensitivity for the characterization of complex chemical and biological samples.
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2 protocols using vnmrs 800 mhz
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NMR Spectroscopy for Biomolecular Characterization
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NMR Spectroscopy of Nucleic Acids
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All NMR experiments were recorded on Agilent Technologies DD2 600 MHz and VNMRS 800 MHz NMR spectrometers at 25°C unless stated otherwise. For suppression of the water signal, the double-pulsed field gradient spin echo (DPFGSE) pulse sequence was used. The translation diffusion coefficients were obtained with the use of pulse field gradient stimulated echo (PFG-STE) pulse sequence. Identification of guanine H1 protons in partially (8%) 13C and 15N site-specific labelled samples was acquired with 1D 15N-edited heteronuclear single quantum correlation (HSQC) experiment. Aromatic protons of adenines and guanines were identified with the use of 2D 13C-edited HSQC experiment. Non-exchangeable proton resonances were assigned using the 2D Nuclear Overhauser Effect SpectroscopY (NOESY) with mixing times (τm) of 80, 150 and 250 ms recorded on NMR samples in 100% D2O. 2D Total Correlation Spectroscopy (TOCSY) with τm of 80 ms, 2D Double Quantum Filtered Correlation SpectroscopY (DQF-COSY) and 2D 1H-31P COSY were used for cross-checking assignment of 2D NOESY spectra. Exchangeable proton resonances were assigned using 2D NOESY experiments with τm of 150 and 250 ms acquired on samples in 90% H2O, 10% D2O.
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