NMR Protocols

Unix/Linux Basics

• Linux basic information and commands

Data Transfer

• Use sftp for data transfer from NMR workstations

Spectrometer operation and probe tuning

• Probe tuning monitored by qtune
• NMR400 BBP/ATB probe tuning procedure
• Reset NMR console if communication is lost
• Variable temperature (VT) experiments on NMR500

Data Processing

• Mnova
• Mnova software license and activation
• Basic 1D processing
• 2D spectrum processing
• Mnova Manual (v12.0.1)
• Mnova Manual (v14.2.0)
• ¹¹B Spectrum: Removing Broad Background Signal with Processing
• Estimate Signal-to-Noise Ratio in vnmrJ and Mnova
• DOSY: Youtube video instruction by Dr Vadim Zorinon on how to process DOSY data with Mnova
• DOSY: Steps to obtain diffusion constants in Mnova
• Removing Broad Background in F19/B11 Spectrum During Processing in Mnova

Printing

• Printing in vnmrJ
• Fixing print quality problems on NMR workstations

General techniques

• Sample volume: detrimental effects of low sample volume on NMR signal
• Kinetics or time-course measurement on Varian instrument
• Time-course or reaction monitoring by NMR on Bruker instrument
• Chemical shift referencing
• Baseline correction for good spectral quality and integration accuracy
• 1H pw90 calibration
• Parameter setting for optimal sensitivity and integration accuracy
• Display saved integral reset points (disable autoprocessing after file loading)
• Varian: Gradient Shimming with Strong H1 Signal
• IconNMR: Running Experiments Without Deuterated Solvents
• Adding separately collected data to create summed spectrum
• Proper procedure to clean NMR tubes (from Wilmad)

1D experiments

• Basic H1 and C13 detection (with H1→C13 NOE)
• {H1}P31 experiment
• {P31}H1 experiment
• Lithium (Li6 and Li7) detection
• H2 (deuterium) detection
• F19 detection
• ¹¹B NMR: Removal of Broad Background Signals
• HOMODEC: identify J-coupled protons
• DPFGSE-gradient NOESY
• 1D and 2D NOESY comparison
• CYCLENOE (NOE difference experiment)
• Impact of CYCLENOE parameter settings
• DEPT: Multiplicity analysis
• DEPTNOE: DEPT and 1H-NOE enhanced 13C spectrum
• Proton T1 relaxation time measurement

2D experiments

All 2D experiments are recommended to be done on Bruker 500 or Varian 600 unless otherwise noted.

Bruker 500

• Time saving with non-uniform sampling (NUS) on Bruker instruments
• Heteronuclear NOESY (HOESY) on Bruker 400
• 2D 13C INADEQUATE Experiment

Varian 600
Please follow the general steps in "Shared steps for all 2D experiment setup". The rest of the instructions for each individual experiment were originally written for NMR500 and have not been fully revised to reflect current hardware configuration.

• Shared steps for all 2D experiment setup on Varian 600
• 2D data processing with nmrPipe/nmrDraw
• 2D data processing with Mnova
• 1H-1H NOESY
• 1H-1H COSY
• 1H-13C HSQC
• 1H-13C HMQC
• 1H-13C HMBC
• 1H-1H TOCSY
• 1H-13C HSQC-TOCSY
• 1H-13C HMQC-TOCSY
• DOSY: examples from 600 MHz spectrometer

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Updated, 03/2021, H. Zhou