Rigor and Reproducibility
Eight steps to Rigorous and Reproducible Experiments in Biomolecular Research at UNC
- If using a core facility, consult with the core staff in the planning stage. Consult with a statistician if you need help developing a Power Analysis to assure that your results will be adequately powered.
- Design your experiment with sufficient controls (rigor) and replicates (reproducibility).
- Assure that ALL of your reagents (antibodies, cell lines, mice) are fully validated (see below).
- Have a clear and detailed protocol (SOP) and data analysis plan. Assure that the protocol is strictly followed or that any deviation is well documented.
- Assure that the staff or students performing the experiment are well trained and understand each step and the importance of performing them precisely.
- Use only well-maintained instrumentation, preferably maintained and operated in a core facility with expert staff (see #1 above).
- Document all steps, reagents, equipment and data analysis methods used in the experiment. Assure that the both the documentation and the data itself are properly stored in a safe data management repository.
- Acknowledge the Cancer Center Support Grant (P30 CA016086) (if applicable), the core (by name), and core staff in publications.
Guide to Rigor and Reproducibility for the Department of Biochemistry and Biophysics Biomolecular Nuclear Magnetic Resonance (NMR) Core Laboratory
- Consult with the core staff in the planning stage. – Stuart Parnham, PhD., Director stitch@email.unc.edu, (919)966-6184.
- NMR sample preparation SOPs can be accessed here, please consult with NMR Core Director if your sample falls outside of these guidelines before using the instruments.
- All New users must attend a training course with the Biomolecular NMR Director before gaining access to the Core facility and its instrumentation.
For more details on sample preparation and more common NMR experiments and applications, please scroll to the bottom of this page to access our SOPs.
Instrument Calibration Standards for all NMR Spectrometers
Temperature calibration:
100% Methanol-d4, 40 mm filling height.
Shim maps:
2 mM Sucrose, 0.5 mM DSS, 2 mM NaN3, 10% D2O, 90% H2O and a 40 mm Filling Height.
Pulse calibrations:
0.1 M 15N-Urea, 0.1 M 13C- CH3OH in 100% DMSO-d6, 40 mm filling height.
48.5 mM TPP in 100% Acetone-d6 for 31P – Sensitivity, 40 mm filling height.
0.05% α, α, α-F3-CC6H5 in 100% C6D6 for 19F – Sensitivity, 40 mm filling height.
0.8 mM 13C 15N – GB-1n 20 mM PO4, 100 mM NaCl, pH 6.5, 0.01% NaN3, 10% D2O, 90% H2O and a 40 mm filling height.
Line-shape:
0.3% CHCl3 in 100% Acetone-d6, 40 mm filling height.
Signal to Noise:
0.1% Ethylbenzine in 100% CDCl3, 40 mm filling height.
Important Protocols
NMR acqisition checklist Protocol