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The Pulse FT NMR Experiment

90º pulse. Experiment. (t). equilibration. detection of signals. Fourier Transform. Data Analysis. Time domain (t). The Pulse FT NMR Experiment. Proteins Have Too Many Signals! 1 H NMR Spectrum of Ubiquitin. ~500 resonances. Resolve resonances by multi-dimensional experiments.

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The Pulse FT NMR Experiment

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  1. 90º pulse Experiment (t) equilibration detection of signals Fourier Transform Data Analysis Time domain (t) The Pulse FT NMR Experiment

  2. Proteins Have Too Many Signals! 1H NMR Spectrum of Ubiquitin ~500 resonances • Resolve resonances by multi-dimensional experiments

  3. [2nd preparation] The 2D NMR Pulse Sequence + 1D + 1D = 2D

  4. 2D NMR: Coupling is the Key 2D detect signals twice (before/after coupling) 90º pulse t1 t2 Transfers between coupled spins Same as 1D experiment t1 t2

  5. t1 t2 The 2D NMR Spectrum Pulse Sequence Spectrum Before mixing Coupled spins After mixing

  6. The Power of 2D NMR:Resolving Overlapping Signals 1D 2 signals overlapped 2D 2 cross peaks resolved

  7. Acronyms For Basic ExperimentsDiffer Only By The Nature Of Mixing Homonuclear Heteronuclear Scalar Coupling COSY HSQC TOCSY Hetero-TOCSY HMQC Multiple Quantum Dipolar Coupling NOESY NOESY-HSQC NOESY-HMQC

  8. t2 t1 t3 Higher Dimensional NMR:Built on the 2D Principle 3D- detect signals 3 times 90º pulse (t3) Same as 1D experiment 3D NMR Pulse Sequence • Experiments are composites  acronyms are composites

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