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How to Get Started on an NMR Project

1. Expression System/Cell Types

  • BL21(DE3)
  • HMS174(DE3)

We typically use pET expression vectors 

2. Appropriate Buffers

  • Tris
  • Hepes
  • Phosphate

If you have questions about a buffer, just give us a call.

3. Sample Conditions

  • Protein Concentration: We recommend a protein concentation of between 0.2mM - 2mM (less than 0.2mM is possible for some experiments if necessary)
  • Sample Volume: Samples need to be 600ul if using a standard NMR tube, or no less than 300ul if using a Shigemi tube
  • D2O: The final NMR sample must contain between 5% and 10% D2O
  • Salt Concentrations: Try to keep this LOW (<100mM total ionic strength)

4. Things to buy:

NMR Core - Protein NMR Data Graphic
The above image illustrates typical preliminary data collected at the start of a protein NMR
project.  The above spectrum takes approximately 15 minutes to collect and tells much about
the feasibility of  a given NMR project.   NMR structural work is carried out on proteins grown with
nutrients labeled with both 15N and 13C.